Figure 5.
TAT-Cx43266-283 blocks autophagy in nutrient-deprived dormant GSCs. Images were acquired by confocal microscopy. (a) Starvation induces autophagy in G166 GSCs, as evidenced by the increased number and intensity staining of LC3 vesicles and decreased number of vesicles and intensity staining of p62. Scale bar: 50 μm. (b) Scheme of the experimental design. G166 GSCs were fixed at the indicated time points, and autophagic flux was assessed by immunofluorescence analysis of LC3 and p62 staining. (c) Representative images of the effect of TAT-Cx43266-283 (50 μM) and CQ (50 μM) on the autophagic flux of starvation-induced dormant G166 GSCs, assessed by LC3 and p62 immunofluorescence. Note the increase in LC3 and p62 vesicles in TAT-Cx43266-283-treated and CQ-treated GSCs, indicative of autophagosome accumulation and impaired autophagic flux. Scale bar: 50 μm (30 min, 4 h), 25 μm (1 h). Similar results were obtained in at least three independent biological replicates. See also related Figure S3 for quantification of these data.