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. Author manuscript; available in PMC: 2015 Jun 23.
Published in final edited form as: Nat Commun. 2014 Dec 23;5:5646. doi: 10.1038/ncomms6646

Figure 5. ALIX regulates injury-triggered accumulation of ESCRT III but not of ALG-2.

Figure 5

(A) Western Blot analysis of ALIX and ALG-2 expression in HeLa cells stably expressing ALIX or empty shRNA vector (Supplementary Fig. 5B shows uncropped blots). Quantification of averaged (B) ALIX, and (C) ALG-2 expression normalized to GAPDH from 3 independent experiments. (D-I) TIRF images and quantification of control and ALIX knockdown cells during the course of repair from laser injury (white arrow) to assess the effect on injury-triggered accumulation of (D, E) ALG2-GFP, (F, G) Chmp4B-GFP, and (H, I) Vps4B-GFP at the site of repair. The plot shows the averaged values for change in the level of respective proteins within the site of repair contained by the dotted box. (J) Western Blot analysis of Vps4B expression in HeLa cells stably expressing Vps4B shRNA or empty shRNA vector (Supplementary Fig. 5B shows uncropped blots). (K) Quantification of averaged Vps4B expression normalized to GAPDH in 3 independent experiments. (L-O) TIRF images and quantification of injury-triggered accumulation of (L, M) ALG2-GFP, and (N, O) Chmp4B-GFP at the site of repair in Vps4B knockdown cells prior to and following repair from laser injury. Injury site is marked by white arrow and the plot shows the averaged values (n=5 cells) for the accumulation of respective proteins. Scale bar represents 10μm and the error bars indicate SE.