Fig. 7: Rab35 regulates actin dynamics.

a Top panels depict change in membrane velocities over time in described conditions. Green represents protraction and red represents retraction of cell membrane. Arrows indicate directionality. The bottom panels are heat maps in which the Red is indicative of retractive movement and green is protractive movement over time. Yellow indicates no change in velocity. b Quantification of cell membrane velocities between indicated groups. Above the dashed line is the protractive velocities and below the dashed line is retractive velocities. n = number of cells. Error bars represent standard deviation, middle bars are the mean. c Western blot of globular (G) and filamentous (F) actin in siRNA (si)-treated groups. d Quantification of the ratio of globular to filamentous actin from blots represented in panel (c). n = number of cells. Error bars represent standard deviation, middle bars are the mean. e Representative images of cells stained for globular and filamentous actin between indicated conditions. f Quantification of the ratio of globular to filamentous actin fluorescent intensities. n = number of cells. Error bars represent standard deviation, middle bars are the mean. g Scanning electron microscopy of filament network between groups. Top panel is the lowest magnification with higher magnifications in panels (i) and (ii). Bottom- cartoon representation of SEM filament network and hypothesized role of Rab35. NS = non-significant. Statistical significance was assessed with an unpaired t-test or a 1-way ANOVA followed by a Dunnett multiple comparisons test. Insets are areas of higher magnification. All experiments were done using human umbilical vein endothelial cells in triplicate.