Videos

Baculovirus actin-based motility drives nuclear envelope disruption and nuclear egress.
Ohkawa T, Welch MD.
Curr Biol 28 2153-2159.e4 (2018)

File OhkawaWelch2018_S1.mp4 Video S1 (Ohkawa-Welch-2018): Actin-Based Motility within the Nucleus and within Protrusions of the Nuclear Envelope. Related to Figure 1. Segment 1: Intra-nuclear actin-based motility. AcMNPV-3Dr nucleocapsids (red) and F-Actin (Lifeact-mCherry; green) in a High Five cell at 29.8 hpi. Video speed is 10 fps; images were captured at 5 s intervals. Scale bar, 10 μm. Segment 2: Circular movements of nucleocapsid and associated actin tails within protrusions of the nuclear envelope. AcMNPV-3Dr nucleocapsid (green), F-actin (Lifeact-tagBFP, blue), and lamin B2 (mCherry-lamin B2, red) in a High Five cell imaged starting at 29.5 hpi. Video speed is 10 fps; images were captured at 5 s intervals. Scale bar, 5 μm. Segment 3: Circular movement of nucleocapsids with associated actin comets at the nuclear periphery. AcMNPV-3Dr nucleocapsids (red) and F-Actin (Lifeact-mCherry; green) in a High Five cell imaged starting at 27.1 hpi. Arrows point to the small donut-shaped actin comet tail structures at the nuclear periphery. Video speed is 10 fps; images were captured at 5 s intervals. Scale bar, 6.7 μm.

File OhkawaWelch2018_S2.mp4 Video S2 (Ohkawa-Welch-2018): Actin-Dependent Viral Disruption of the Nuclear Envelope. Related to Figure 3. Segment 1: Disruption of the nuclear envelope, as visualized by localized dissolution of the nuclear lamina. Left panel: F-actin (Lifeact-tagBFP, green), and lamin B2 (mCherry-lamin B2, red) in a High Five cell imaged starting at 24 hpi. The white box highlights an area of the nuclear envelope in which loss of the mCherry-lamin B2 signal is accompanied by a transient burst of Lifeact-tagBFP. A magnified image is in the lower left corner. Video speed is 10 fps; images were captured at 10 s intervals. Scale bar, 10 μm. Right panel: A second example of nuclear envelope disruption, as visualized by localized dissolution of the nuclear envelope. F-actin (Lifeact-tagBFP, green), and lamin B2 (mCherry-lamin B2, red) in a High Five cell imaged starting at 28 hpi. The white box highlights the area of the nuclear envelope in which loss of the mCherry-lamin B2 signal is accompanied by a transient burst of the Lifeact-tagBFP signal as well as the appearance of an actin comet tail as a nucleocapsid escapes from the nucleus. A magnified image is in the lower right corner. Video speed is 10 fps; images were captured at 10 s intervals. Arrow points to the protrusion in the nuclear envelope. Scale bar, 20 μm. Segment 2: Disruption of the nuclear envelope, as visualized by localized dissolution of the nuclear lamina. Actin (EGFP-actin; red) and SUN2-3xmCherry (green) in an AcMNPV-WOBpos infected High Five cell imaged starting at 25.7 hpi. Video speed is 10 fps; images were captured at 10 s intervals. Scale bar, 5 μm.

File OhkawaWelch2018_S3.mp4 Video S3 (Ohkawa-Welch-2018): Loss of NE Integrity in Infected Cells as Visualized by Leakage of NLS-GFP from the Nucleus. Related to Figures 3 and 4. Loss of nuclear NLS-GFP signal during late stage AcMNPV infection. Top panels: NLS-GFP in a High Five cell infected with AcMNPV-3mCq, imaged starting at 11.2 hpi; the top left panel shows NLS-GFP signal while the top right panel shows VP39-3xmCherry. Video speed is 10 fps; images were captured at 15 min intervals. Time in hr:min. Scale bar, 10 μm. Bottom panels: retention of nuclear NLS-GFP in an uninfected High Five cell; the bottom left panel shows NLS-GFP signal while the bottom right panel shows VP39-3xmCherry. Video speed is 10 fps; images were captured at 15 min intervals. Time in hr:min. Scale bar, 10 μm.

File OhkawaWelch2018_S4.mp4 Video S4 (Ohkawa-Welch-2018): Blocking Nuclear Pores or CRM1/Exportin 1 Export Does Not Affect AcMNPV Egress from the Nucleus. Related to Figure 3. Segment 1: WGA treatment does not prevent AcMNPV egress from the nucleus. Right panel: a High Five cell infected with AcMNPV WOBpos and microinjected with WGA-TRITC is imaged. The WGA-TRITC signal is shown, outlining the nucleus at the onset of imaging. The cell is transfected with pACT-GFP-actin, although EGFP-actin is not shown in this video. Video speed is 10 fps; images were captured at 15 min intervals. Time in hr:min. Scale bar, 10 μm. Left panel: the same High Five cell is shown, with the EGFP-actin signal visualized. Cytoplasmic actin-based motility is visible by 19:40 hpi. Video speed is 10 fps; images were captured at 15 min intervals. Video frames are paused to indicate cytoplasmic actin comet tails with arrows. Time in hr:min. Scale bar, 10 μm. Segment 2: Leptomycin B treatment of infected cells does not prevent AcMNPV egress from the nucleus. High Five cell expressing EGFP-actin, infected with AcMNPV-3mC, with Leptomycin B added at 15 hpi. EGFP-actin signal is shown, with a burst of nuclear actin comet tail activity at 17:08 hpi, and cytoplasmic actin comet tails visible by 18:18 hpi. Video speed is 10 fps; images were captured at 10 min intervals. Video frames are paused to indicate cytoplasmic actin comet tails with arrows. Time in hr:min. Scale bar, 10 μm

File OhkawaWelch2018_S5.mp4 Video S5 (Ohkawa-Welch-2018): AcMNPV Undergoes Actin-Based Motility in the Cytoplasm following Nuclear Escape. Related to Figure 3. AcMNPV uses actin-based motility in the cytoplasm after escape from the nucleus. AcMNPV-3Dr nucleocapsids (red) and F-Actin (Lifeact-mCherry; green) in a High Five cell imaged starting at 27.6 hpi. Video speed is 10 fps; images were captured at 8 s intervals. Scale bar, 10 μm.

Virulent Burkholderia species mimic host actin polymerases to drive actin-based motility.
Benanti EL, Nguyen CM, Welch MD.
Cell 161 348-360 (2015)

File benantinguyen2015_1(1).mp4 Video 1 (Benanti-Nguyen-2015): BmBimA processively binds and elongates filament barbed ends. TIRF microscopy of Alexa Fluor 488-labeled BmBimA (10 nM) elongation of rhodamine-labeled actin filaments. Rhodamine-actin is magenta, and BmBimA is green. Scale bar, 3 μm.

File benantinguyen2015_2.mp4 Video 2 (Benanti-Nguyen-2015): BpBimA processively binds and elongates filament barbed ends. TIRF microscopy of Alexa Fluor 488-labeled BpBimA (100 pM) elongation of rhodamine-labeled actin filaments. Rhodamine-actin is magenta, and BpBimA is green. Scale bar, 3 μm.

File benantinguyen2015_3(1).mp4 Video 3 (Benanti-Nguyen-2015): BmBimA elongates two filaments simultaneously. TIRF microscopy of Alexa Fluor 488-labeled BmBimA (10 nM) binding and elongating two actin filaments. Rhodamine-actin is magenta, and BmBimA is green. Scale bars, 3 μm.

File benantinguyen2015_4(1).mp4 Video 4 (Benanti-Nguyen-2015): BmBimA elongates three filaments simultaneously. TIRF microscopy of Alexa Fluor 488-labeled BmBimA (10 nM) binding and elongating three actin filaments. Rhodamine-actin is magenta, and BmBimA is green. Scale bars, 3 μm.

File benantinguyen2015_5(1).mp4 Video 5 (Benanti-Nguyen-2015): BpBimA elongates two filaments simultaneously. TIRF microscopy of Alexa Fluor 488-labeled BpBimA (100 pM) binding and elongating two actin filaments. Rhodamine-actin is magenta, and BpBimA is green. Scale bars, 3 μm.

File benantinguyen2015_6(1).mp4 Video 6 (Benanti-Nguyen-2015): BpBimA elongates three filaments simultaneously. TIRF microscopy of Alexa Fluor 488-labeled BpBimA (100 pM) binding and elongating three actin filaments. The initial two-filament bundle and subsequent three-filament bundle are indicated by arrowheads. Rhodamine-actin is magenta, and BpBimA is green. Scale bars, 3 μm.

File benantinguyen2015_7(1).mp4 Video 7 (Benanti-Nguyen-2015): BimA orthologs mediate distinct modes of actin-based motility in B. thailandensis. From left to right, BtBimA-, BpBimA-, or BmBimA-expressing B. thailandensis infections of Cos7 Lifeact-EGFP cells. Bacteria also constitutively express RFP and are shown in magenta. Lifeact-EGFP is shown in green. Scale bars, 10 μm.

Rickettsia actin-based motility occurs in distinct phases mediated by different actin nucleators.
Reed SC, Lamason RL, Risca VI, Abernathy E, Welch MD.
Curr Biol 24 98-103 (2014)

File reedlamason2014_1.mp4 Video 1 (Reed-Lamason-2014): Live HMEC-1 cells transfected with EGFP-Lifeact (green) and infected for 20 min with R. parkeri expressing mCherry (red). Frames were imaged every 5 s; movie plays at 10 frames/s (50x actual speed). Time stamp is in min:s.

File reedlamason2014_2.mp4 Video 2 (Reed-Lamason-2014): Live HMEC-1 cells transfected with EGFP-Lifeact (green) and infected for 48 hr with R. parkeri expressing mCherry (red). Frames were imaged every 5 s; movie plays at 10 frames/s (50x actual speed). Time stamp is in min:s.

File reedlamason2014_3.mp4 Video 3 (Reed-Lamason-2014): Live HMEC-1 cells stably expressing mCherry-Lifeact (green) and infected for 9 hr with L. monocytogenes 10403S expressing EGFP (red). Frames were imaged every 5 s; movie plays at 10 frames/s (50x actual speed). Time stamp is in min:s.

Regulation of integrin trafficking, cell adhesion, and cell migration by WASH and the Arp2/3 complex.
Duleh SN, Welch MD.
Cytoskeleton 69 1047-1058 (2012)

File dulehwelch2010_1.mp4 Video 1 (Duleh-Welch-2012): WASH and F-actin dynamics on subdomains of Rab5-Q79L endosomes. NIH3T3 cells transfected with pGFP-WASH (green), pLifeact-BFP (blue) to mark F-actin, and pDsRed-Rab5-Q79L (red) were imaged on a spinning disc confocal microscope.

File dulehwelch2012_2.mp4 Video 2 (Duleh-Welch-2012): WASH and F-actin dynamics on subdomains of an enlarged endosome. Magnified from Movie 1, corresponding to the inset of Figure 1A.

Rickettsia parkeri invasion of diverse host cells involves an Arp2/3 complex, WAVE complex and Rho-family GTPase-dependent pathway.
Reed SC, Serio AW, Welch MD.
Cell Microbiol 14 529-545 (2012)

File reedserio2012_1.mp4 Video 1 (Reed-Serio-2012): Actin recruitment around a group of invading Rickettsia. R. parkeri GFP-3 (red) is shown in a Cos7 cell expressing LifeAct-mCherry (green). Images were captured every 10 s starting approximately 23 min post infection and are displayed at 10 frames per second. Bar, 5 μm.

File reedserio2012_2.mp4 Video 2 (Reed-Serio-2012): Two internalizing bacteria associated with actin. R. parkeri GFP-3 (red) is shown in a Cos7 cell expressing LifeAct-mCherry (green). Images were captured every 10 s starting approximately 30 min post infection and are displayed at 10 frames per second. Bar, 5 μm.

File reedserio2012_3.mp4 Video 3 (Reed-Serio-2012): Internalization of a bacterium surrounded by mCherry-PAK-PBD. R. parkeri GFP-3 (red) is shown in a Cos7 cell expressing mCherry-PAK-PBD (green). Images were captured every 10 s starting approximately 11 min post infection and are displayed at 10 frames per second. Bar, 5 μm.

File reedserio2012_4.mp4 Video 4 (Reed-Serio-2012): Internalization of two individual bacteria associated with mCherry-PAK-PBD. R. parkeri GFP-3 (red) is shown in a Cos7 cell expressing LifeAct-mCherry (green). Images were captured every 10 s starting approximately 30 min post infection and are displayed at 10 frames per second. Bar, 5 μm.

The actin nucleation factor JMY is a negative regulator of neuritogenesis.
Firat-Karalar EN, Hsiue PP, Welch MD.
Mol Biol Cell 22 4563-4574 (2011)

Movie icon firat-karalarhsiue2011_1.mov Video 1 (Firat-Karalarhsiue-2011): Polymerization of rhodamine-actin and actin-based motility of beads coated with JMY-WWWCA in Xenopus laevis extract, imaged by epifluorescence microscopy.

An actin-filament-binding interface on the Arp2/3 complex is critical for nucleation and branch stability.
Goley ED, Rammohan A, Znameroski EA, Firat-Karalar EN, Sept D, Welch MD.
Proc Natl Acad Sci U S A 107 8159-8164 (2010)

File goleyrammohan2010_1.mp4 Video 1 (Goley-Rammohan-2010): Three-dimensional view comparing models of the interaction between ARPC2/ARPC4 and F actin. The position of ARPC2/ARPC4 derived from protein-protein docking simulations is depicted in pink and cyan, whereas the position of these subunits suggested by Rouiller et al. (2008, J Cell Biol 180:887-895) is shown in yellow. The actin filament is depicted in white.

Actin-based motility drives baculovirus transit to the nucleus and cell surface.
Ohkawa T, Volkman LE, Welch MD.
J Cell Biol 190 187-195 (2010)

File ohkawavolkman2010_1.mp4 Video 1 (Ohkawa-Volkman-2010): Actin-based motility of AcMNPV 3mC virus (red) is shown in a High Five cell expressing EGFP-actin (green).

File ohkawavolkman2010_2.mp4 Video 2 (Ohkawa-Volkman-2010): Comparison of the motility of AcMNPV 3mC WT (red; left) and I358A mutant (red; right) in High Five cells expressing EGFP-actin (green).

File ohkawavolkman2010_3.mp4 Video 3 (Ohkawa-Volkman-2010): Actin-based motility of AcMNPV 3mC I358A virus (red) in a High Five cell expressing EGFP-actin (green).

File ohkawavolkman2010_4.mp4 Video 4 (Ohkawa-Volkman-2010): Comparison of the movement tracks of AcMNPV WT and I358A mutant.

File ohkawavolkman2010_5.mp4 Video 5 (Ohkawa-Volkman-2010): AcMNPV 3mC nucleocapsids (red) are shown colliding with the nucleus in High Five cells expressing EGFP- actin (green). The yellow circle highlights a virus colliding with the nucleus, although other collisions are also apparent.

File ohkawavolkman2010_6.mp4 Video 6 (Ohkawa-Volkman-2010): AcMNPV 3mC nucleocapsids (red) are shown colliding with the nucleus in High Five cells expressing EGFP- actin (green).

File ohkawavolkman2010_7.mp4 Video 7 (Ohkawa-Volkman-2010): EGFP-actin is shown in High Five cells 5 min after infection. The yellow circle highlights a virus colliding with the nucleus, although other collisions are also apparent. After collisions, virus-associated actin comet tails persist, resembling corkscrews that radiate from the nuclear periphery.

File ohkawavolkman2010_8.mp4 Video 8 (Ohkawa-Volkman-2010): AcMNPV 3mC nucleocapsids (red) are shown separating from their actin comets and entering the nucleus in High Five cells expressing EGFP-actin (green). After nucleocapsid entry into the nucleus, actin polymerization ceases.

File ohkawavolkman2010_9.mp4 Video 9 (Ohkawa-Volkman-2010): An AcMNPV 3mC I358A nucleocapsid (red) is shown with a virus-associated actin corkscrew structure radiating from the nuclear periphery in High Five cells expressing EGFP-actin (green).

File ohkawavolkman2010_10.mp4 Video 10 (Ohkawa-Volkman-2010): AcMNPV 3mC virus (red) is shown moving into surface spikes in a High Five cell expressing EGFP-actin (green). This video is a magnified and cropped view from Video 1.

WASH and the Arp2/3 complex regulate endosome shape and trafficking.
Duleh SN, Welch MD.
Cytoskeleton 67 193-206 (2010)

File dulehwelch2010_1.mp4 Video 1 (Duleh-Welch-2010): WASH (green) is asymmetrically distributed on EEA1-positive endosomes (red) as visualized by deconvolution microscopy and 3D reconstruction.

Rickettsia Sca2 is a bacterial formin-like mediator of actin-based motility.
Haglund CM, Choe JE, Skau CT, Kovar DR, Welch MD.
Nat Cell Biol 12 1057-1063 (2010)

File haglundchoe2010_1 .mp4 Video 1 (Haglund-Choe-2010): Polymerization of actin filaments under control conditions, imaged by TIRF microscopy. A red dot marks the pointed end of a control filament (c), and a red open arrowhead marks the growing barbed end.

File haglundchoe2010_2.mp4 Video 2 (Haglund-Choe-2010): Polymerization of actin filaments in the presence of GST- Sca2, imaged by TIRF microscopy. Green filled arrowheads mark Sca2-associated filaments (s). A red dot marks the pointed end of a control filament (c), and a red open arrowhead marks the growing barbed end.

File haglundchoe2010_3.mp4 Video 3 (Haglund-Choe-2010): Polymerization of actin filaments in the presence of GST-mDia2(FH1FH2), imaged by TIRF microscopy. A red dot marks the pointed end of a control filament (c), and a red open arrowhead marks the growing barbed end. A blue dot (m) marks the pointed end of an mDia2-associated filament, and a blue open arrowhead marks the growing barbed end.

File haglundchoe2010_4.mp4 Video 4 (Haglund-Choe-2010): Polymerization of actin filaments in the presence of GST-mDia2(FH1FH2) and profilin, imaged by TIRF microscopy. A blue dot marks the pointed end of an mDia2-bound filament (m), and a blue open arrowhead marks the growing barbed end. A red dot marks the pointed end of a control filament (c), and a red open arrowhead marks the growing barbed end.

File haglundchoe2010_5.mp4 Video 5 (Haglund-Choe-2010): Polymerization of actin filaments in the presence of profilin, imaged by TIRF microscopy. A red dot marks the pointed end of a control filament (c), and a red open arrowhead marks the growing barbed end.

File haglundchoe2010_6.mp4 Video 6 (Haglund-Choe-2010): Polymerization of actin filaments in the presence of profilin and GST-Sca2, imaged by TIRF microscopy. A red dot marks the pointed end of a control filament (c), and a red open arrowhead marks the growing barbed end. A green dot marks the pointed end of a Sca2-bound filament (s), and a green open arrowhead marks the growing barbed end.

File haglundchoe2010_7.mp4 Video 7 (Haglund-Choe-2010): Polymerization of actin filaments in the presence of immobilized GST-mDia2(FH1FH2) and profilin, imaged by TIRF microscopy. A blue dot marks the pointed end of an mDia2-bound filament (m), and a blue open circle marks the growing barbed end.

File haglundchoe2010_8.mp4 Video 8 (Haglund-Choe-2010): Polymerization of actin filaments in the presence of immobilized GST-Sca2 and profilin, imaged by TIRF microscopy. A green dot marks the pointed end of a Sca2-bound filament (s), and a green open circle marks the growing barbed end.

File haglundchoe2010_9.mp4 Video 9 (Haglund-Choe-2010): Polymerization of rhodamine-actin on beads coated with GST-Sca2 in Xenopus laevis extract, imaged by epifluorescence microscopy (top) and phase-contrast microscopy (bottom).

Defining a core set of actin cytoskeletal proteins critical for actin-based motility of Rickettsia.
Serio AW, Jeng RL, Haglund CM, Reed SC, Welch MD.
Cell Host Microbe 7 388-398 (2010)

File seriojeng2010_1.mp4 Video 1 (Serio-Jeng-2010): Actin labeled with Lifeact-GFP in Rickettsia parkeri infected S2R+ cells not treated with dsRNA (left) or treated with dsRNA targeting capping protein (right).

File seriojeng2010_2.mp4 Video 2 (Serio-Jeng-2010): Actin labeled with Lifeact-GFP in Rickettsia parkeri infected S2R+ cells treated with dsRNA targeting cofilin (left) or fimbrin (right).

File seriojeng2010_3.mp4 Video 3 (Serio-Jeng-2010): Actin labeled with Lifeact-GFP in Rickettsia parkeri infected S2R+ cells treated with dsRNA targeting profilin, showing a short tail phenotype (left) or short actin spikes (right).

File seriojeng2010_4.mp4 Video 4 (Serio-Jeng-2010): Plastin-GFP in COS7 cells infected with Rickettsia parkeri (left) or Listeria monocytogenes (right).

File seriojeng2010_5.mp4 Video 5 (Serio-Jeng-2010): Profilin-GFP in COS7 cells infected with Rickettsia parkeri (left) or Listeria monocytogenes (right).

File seriojeng2010_6.mp4 Video 6 (Serio-Jeng-2010): GFP-capping protein in COS7 cells infected with Rickettsia parkeri (left) or Listeria monocytogenes (right).

File seriojeng2010_7.mp4 Video 7 (Serio-Jeng-2010): GFP-cofilin in COS7 cells infected with Rickettsia parkeri (left) or Listeria monocytogenes (right).

File seriojeng2010_8.mp4 Video 8 (Serio-Jeng-2010): Phase contrast images (top) or GFP fluorescence (bottom) in COS7 cells infected with Rickettsia parkeri (left) or Listeria monocytogenes (right).

File seriojeng2010_9.mp4 Video 9 (Serio-Jeng-2010): Actin labeled with Lifeact-GFP in a Rickettsia parkeri infected COS7 cell transfected with a control non- specific siRNA.

File seriojeng2010_10.mp4 Video 10 (Serio-Jeng-2010): Actin labeled with Lifeact-GFP in a Rickettsia parkeri infected COS7 cell depleted of T-plastin by RNAi.

File seriojeng2010_11.mp4 Video 11 (Serio-Jeng-2010): Actin labeled with Lifeact-GFP in a Rickettsia parkeri infected COS7 cell depleted of profilin 1 by RNAi.

WHAMM is an Arp2/3 complex activator that binds microtubules and functions in ER to Golgi transport.
Campellone KG, Webb NJ, Znameroski EA, Welch MD.
Cell 134 148-161 (2008)

File campellonewebb2008_1.mp4 Video 1 (Campellone-Webb-2008): Dynamics of GFP-WHAMM-associated membranes in a Cos7 cell over a 10 min timecourse.

File campellonewebb2008_2.mp4 Video 2 (Campellone-Webb-2008): Dynamics of GFP-WHAMM-associated membranes in a Cos7 cell over a 6 min timecourse.

File campellonewebb2008_3.mp4 Video 3 (Campellone-Webb-2008): Dynamics of GFP-WHAMM-associated membranes in an NIH 3T3 cell in the presence of nocodazole. The drug was added 3 min into a 9 min time series.

File campellonewebb2008_4.mp4 Video 4 (Campellone-Webb-2008): Dynamics of GFP-WHAMM-associated membranes in an NIH 3T3 cell in the presence of cytochalasin D. The drug was added 3 min into a 9 min time series.

File campellonewebb2008_5.mp4 Video 5 (Campellone-Webb-2008): Dynamics of GFP-WHAMM-associated membranes in an NIH 3T3 cell in the presence of latrunculin A. The drug was added 3 min into a 9 min time series.

File campellonewebb2008_6.mp4 Video 6 (Campellone-Webb-2008): Dynamics of GFP-WAMM(W807A)-associated tubules in an NIH 3T3 cell over a 6 min timecourse.

Dynamic nuclear actin assembly by Arp2/3 complex and a baculovirus WASP-like protein.
Goley ED, Ohkawa T, Mancuso J, Woodruff JB, D'Alessio JA, Cande WZ, Volkman LE, Welch MD.
Science 314 464-467 (2006)

File goleyohkawa2006_1.mp4 Video 1 (Goley-Ohkawa-2006): Actin localization and polymerization in infected TN-368 cells expressing EGFP-actin, viewed from 14:30 to 21:42 hours post infection (hpi). Latrunculin A is added at approximately 20 hpi, causing depolymerization of the discrete nuclear F-actin structures.

File goleyohkawa2006_2.mp4 Video 2 (Goley-Ohkawa-2006): Actin localization and polymerization in infected TN-368 cells expressing EGFP-actin, viewed from 18:45 to 22:05 hours post infection.

File goleyohkawa2006_3.mp4 Video 3 (Goley-Ohkawa-2006): Actin localization and polymerization in infected TN-368 cells expressing mCherry-actin, viewed from 5:44 to 22:40 hours post infection.

File goleyohkawa2006_4.mp4 Video 4 (Goley-Ohkawa-2006): Fluorescence recovery after photobleaching a region of the nucleus in an infected TN-368 cell expressing EGFP-actin. The circle marks the photobleached region.

File goleyohkawa2006_5.mp4 Video 5 (Goley-Ohkawa-2006): Fluorescence loss in photobleaching of nuclear GFP-actin in a representative infected TN-368 cell that has been treated with latrunculin A.

Plasma membrane organization is essential for balancing competing pseudopod- and uropod-promoting signals during neutrophil polarization and migration.
Bodin S, Welch MD.
Mol Biol Cell 16 5773-5783 (2005)

File bodinwelch2005_1.mp4 Video 1 (Bodin-Welch-2005): Inhibition of the fMLP-induced chemotaxis response in cholesterol-depleted cells. Control cell (left) or MβCD-treated cell (right) were visualized using DIC microscopy.

File bodinwelch2005_2.mp4 Video 2 (Bodin-Welch-2005): Cholesterol-depletion increases the chemoattractant sensitivity of the rear edge. Cell was visualized using DIC microscopy.

File bodinwelch2005_3.mp4 Video 3 (Bodin-Welch-2005): Cholesterol depletion does not inhibit the fMLP-induced formation of a retracting uropod in pertussis toxin- treated cells. Pertussis-toxin (PTX) treated cell (left) and a cell treated with both PTX and MβCD (right), visualized using DIC microscopy.

File bodinwelch2005_4.mp4 Video 4 (Bodin-Welch-2005): Cholesterol is required to restrict D3-PI synthesis to the pseudopod. Control cell (Video 4) and cholesterol-depleted cells (Videos 5 and 6) expressing PH-Akt-GFP, left panels DIC microscopy, right panels PH- Akt-GFP fluorescence.

File bodinwelch2005_5(1).mp4 Video 5 (Bodin-Welch-2005): Cholesterol is required to restrict D3-PI synthesis to the pseudopod. Control cell (Video 4) and cholesterol-depleted cells (Videos 5 and 6) expressing PH-Akt-GFP, left panels DIC microscopy, right panels PH- Akt-GFP fluorescence.

File bodinwelch2005_6(1).mp4 Video 6 (Bodin-Welch-2005): Cholesterol is required to restrict D3-PI synthesis to the pseudopod. Control cell (Video 4) and cholesterol-depleted cells (Videos 5 and 6) expressing PH-Akt-GFP, left panels DIC microscopy, right panels PH- Akt-GFP fluorescence.

File bodinwelch2005_7.mp4 Video 7 (Bodin-Welch-2005): Distribution of D3-PIs in a control cell during an abrupt reversion of the fMLP gradient visualized by PH-Akt- GFP fluorescence.

File bodinwelch2005_8.mp4 Video 8 (Bodin-Welch-2005): Distribution of D3-PIs in a cholesterol- depleted cell during an abrupt reversion of the fMLP-gradient visualized by PH-Akt-GFP fluorescence.

File bodinwelch2005_9.mp4 Video 9 (Bodin-Welch-2005): Cholesterol depletion induces pseudopod formation in a PI3-K- and Gi- dependent manner. Cells expressing PH- Akt-GFP were exposed to a gradient of MbCD diffusing from a micropipette. Cells respond by emitting a pseudopod facing the pipette tip (top panel), accompanied by a modest membrane relocation of PH-Akt-GFP (indicated by white arrow on frame 100 s). This response is prevented in cells pre-treated by pertussis-toxin (middle panel), or wortmannin (lower panel).

A Rickettsia WASP-like protein activates the Arp2/3 complex and mediates actin-based motility.
Jeng RL, Goley ED, D'Alessio JA, Chaga OY, Svitkina TM, Borisy GG, Heinzen RA, Welch MD.
Cell Microbiol 6 761-769 (2004)

File jenggoley2004_1 (1).mp4 Video 1 (Jeng-Goley-2004): Movie of actin structures assembled by RickA-coated beads in Xenopus egg extract that was supplemented with rhodamine-labelled actin and visualized by fluorescence microscopy.