Figure 5. NBR1 facilitates cell migration and FA turnover. (A) Representative phase-contrast microscopy images at time of wounding (0 h) and 6 h after wounding for cells expressing control siRNA (CTRL) or siRNA against NDP52, OPTN, p62, or NBR1. Dashed yellow lines highlight wound boundaries. Bars, 100 µm. (B) Quantification of wound closure over 6 h by cells expressing indicated siRNAs. The decrease in wound width was determined by subtracting the final width at 6 h from the initial width at 0 h. Bar graph shows mean + SEM, representing n = 8 wounds for siCTRL, n = 8 wounds for siNDP52, n = 8 wounds for siOPTN, n = 6 wounds for sip62, n = 6 wounds for siNBR1, and n = 5 wounds for mock (no siRNA) pooled from four independent experiments. P-values calculated using one-way analysis of variance followed by Tukey post-hoc test. n.s., not significant. (C) Representative phase-contrast microscopy images of cells expressing shCTRL or shNBR1 at time of wounding (0 h) and at 5 h after wounding. Dashed yellow lines highlight wound boundaries. Bars, 100 µm. (D) Quantification of wound closure over 5 h by shCTRL and shNBR1 cells. Bar graph shows mean + SEM, representing n = 12 wounds for shCTRL and n = 15 wounds for shNBR1 pooled from three independent experiments. P-value determined using unpaired t test. (E) Spinning disk confocal microscopy time-lapse sequences of cells expressing paxillin-mCherry (black) to monitor FA dynamics. Left panels show representative cells expressing shCTRL (top) or shNBR1 (bottom). Image sequences of boxed regions on the right have been rotated such that the cell edge with dynamic FAs is moving upward vertically. Elapsed time (min) in top left of images. Bars, 5 µm. Insets are magnified twofold. These images correspond to Videos 7 and 8. (F) Quantification of FA assembly rate constants (left), disassembly rate constants (middle), and lifetime (right) for FAs in shCTRL or shNBR1 cells. Data presented as median (line), first and third quartile (box), and whiskers extend to ±1.5 times the interquartile range. Individual data points outside of this range are shown. n = 53 FAs for shCTRL and n = 58 FAs for shNBR1, pooled from three independent experiments. P-value calculated using a nonparametric Mann-Whitney test.
Figure 5.

NBR1 facilitates cell migration and FA turnover. (A) Representative phase-contrast microscopy images at time of wounding (0 h) and 6 h after wounding for cells expressing control siRNA (CTRL) or siRNA against NDP52, OPTN, p62, or NBR1. Dashed yellow lines highlight wound boundaries. Bars, 100 µm. (B) Quantification of wound closure over 6 h by cells expressing indicated siRNAs. The decrease in wound width was determined by subtracting the final width at 6 h from the initial width at 0 h. Bar graph shows mean + SEM, representing n = 8 wounds for siCTRL, n = 8 wounds for siNDP52, n = 8 wounds for siOPTN, n = 6 wounds for sip62, n = 6 wounds for siNBR1, and n = 5 wounds for mock (no siRNA) pooled from four independent experiments. P-values calculated using one-way analysis of variance followed by Tukey post-hoc test. n.s., not significant. (C) Representative phase-contrast microscopy images of cells expressing shCTRL or shNBR1 at time of wounding (0 h) and at 5 h after wounding. Dashed yellow lines highlight wound boundaries. Bars, 100 µm. (D) Quantification of wound closure over 5 h by shCTRL and shNBR1 cells. Bar graph shows mean + SEM, representing n = 12 wounds for shCTRL and n = 15 wounds for shNBR1 pooled from three independent experiments. P-value determined using unpaired t test. (E) Spinning disk confocal microscopy time-lapse sequences of cells expressing paxillin-mCherry (black) to monitor FA dynamics. Left panels show representative cells expressing shCTRL (top) or shNBR1 (bottom). Image sequences of boxed regions on the right have been rotated such that the cell edge with dynamic FAs is moving upward vertically. Elapsed time (min) in top left of images. Bars, 5 µm. Insets are magnified twofold. These images correspond to Videos 7 and 8. (F) Quantification of FA assembly rate constants (left), disassembly rate constants (middle), and lifetime (right) for FAs in shCTRL or shNBR1 cells. Data presented as median (line), first and third quartile (box), and whiskers extend to ±1.5 times the interquartile range. Individual data points outside of this range are shown. n = 53 FAs for shCTRL and n = 58 FAs for shNBR1, pooled from three independent experiments. P-value calculated using a nonparametric Mann-Whitney test.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal