Fig. 7: Involvement of HSPB8-induced autophagy in BLM and A375 cell proliferation.

a, b BLM cells were pre-treated with or without 1 mM 3-MA for 1 h before HSPB8 overexpression (48 h). WB analysis of LC3 and RAS protein were carried out (a) and the effect on cell proliferation was evaluated by cell count (b). Data are mean ± SD of four independent biological samples (N = 4). Each experiment was repeated three times. Statistical analysis was performed using One-way ANOVA followed by Bonferroni post-hoc test; (*p < 0.05). c WB analysis of ATG5 and LC3 in BLM cells transfected with 20 nM negative control siRNA (NC) or ATG5 siRNA for 72 h and HSPB8 overexpression (48 h). d Effect of ATG5 siRNA on proliferation of BLM cells overexpressing HSPB8. Data are mean ± SD of four independent biological samples (N = 4). Each experiment was repeated three times. Statistical analysis was performed using One-way ANOVA followed by Bonferroni post-hoc test; (*p < 0.05; **p < 0.01). e, f A375 cells were pre-treated with or without 1 mM 3-MA for 1 h before HSPB8 overexpression (48 h). At the end of the treatment WB analysis of LC3 and RAS protein were carried out (e) and the effect on cell proliferation was evaluated by cell count (f). Four independent biological samples for each condition were analyzed (N = 4), bar graph represents the mean relative cell viability ± SD. Each experiment was repeated three times. Statistical analysis was performed using One-way ANOVA followed by Bonferroni post-hoc test (*p < 0.05, **p < 0.01). g WB analysis of ATG5 and LC3 in A375 cells transfected with 20 nM negative control siRNA (NC) or ATG5 siRNA for 72 h and HSPB8 overexpression (48 h). h Effect of ATG5 siRNA on cell proliferation of A375 cells overexpressing HSPB8. Data are mean ± SD of four independent biological samples (N = 4). Each experiment was repeated three times. Statistical analysis was performed using One-way ANOVA followed by Bonferroni post-hoc test; (*p < 0.05; **p < 0.01).