Fig. 2.
(A) AG11395 cells were transfected with empty vector (EGFPC1) or full length WRN plasmid and go for ER stress for 8 h. Total cell lysate was immunoblotted with anti-LC3B, anti-Beclin-1, anti-Atg5 antibodies. Here β-Actin was used as a loading control. (B–D) Bands intensity were measured and graphically represent the ratio of LC3B-II and LC3B-I, Beclin-1 and Atg5 at different time point respectively. (E) AG05229 cells were tranfected with full length WRN plasmid and go for ER stress for 8 h. Total cell lysate was immunoblotted with anti-LC3B, anti-Beclin-1, anti-Atg5 antibodies. Here β-Actin was used as a loading control. (F) Graphically represent the ratio of LC3B-II and LC3B-I. Normal (GM00637) and WS (AG11395) cells were transfected with pBABE-puro mCherry-EGFP-LC3 B and ER stressed for 8 h. (G) mCherry and EGFP signal were observed under fluorescence microscopy. Atleast 450 cells were examined for different conditions. (H) Graphical representation of autophagic flux of all cell lines. 3 western blots from 3 separate sets of samples were used. *** p < 0.0005, mean ± SD, n=3.