Fig. 5

mTOR pathway was activated, autophagy was inhibited after HMGB1 knockdown and the enhanced sensitivity effect of HMGB1 knockdown was blocked by CQ (an autophagy inhibitor). a: Western blotting was performed to analyze changes in the mTOR pathway after HMGB1 knockdown; the phosphorylation of p70S6K (substrate of the mTORC1 complex) and AKT-ser473 (substrate of the mTORC2 complex) were both increased; b: Densitometric analysis of the phosphorylation of mTOR, p70S6K and AKT; c: Representative results showing the fluorescence intensities in RPMI8226 after Dex (100 μM) exposure using the Cyto-ID Autophagy Detection Kit with flow cytometry, control shRNA without Dex (red line), HMGB1 shRNA without Dex (blue line), control shRNA with Dex (orange line), and HMGB1 shRNA with Dex (green line); d: Histograms showing the analysis results of the MM cell lines(RPMI8226, CAG) autophagy fluorescence intensities induced by Dex after HMGB1 knockdow by Cyto-ID Autophagy Detection Kit n (n = 3); e: Western blotting was performed to detect the Dex-induced expression of LC3A/B-I/II after HMGB1 knockdown and the histograms showing the densitometric analysis of LC3A/B-II; f: Histograms showing the analysis results of RPMI8226 (transfection with control shRNA or HMGB1 shRNA) apoptosis induced by Dex (100 μM) in the presence of CQ (1 μM, an autophagy inhibitor) (*P<0.05,**P<0.01,***P<0.001)