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. 2021 Nov 10;10(11):3118. doi: 10.3390/cells10113118

Figure 2.

Figure 2

Course of copper-induced autophagy in HepG2 WT and HepG2 ATP7B−/− cells. Western blot analysis of LC3B-II/LC3B-I ratio of copper-treated HepG2 WT (ai) and HepG2 ATP7B−/− (aii) cells. (b) Quantification of western blot band intensity of LC3B-II/LC3B-I ratio. Autophagosome-lysosome fusion analysis in the HepG2 WT (ci) and HepG2 ATP7B−/− cells (cii) under starvation (EBSS) and with copper added. The tandem GFP-RFP-LC3B was used as the indicator for autophagosome-lysosome fusion. Red puncta indicated fused autophagosome with lysosome and the yellow puncta indicated unfused autophagosome. (d) Analysis of red and yellow puncta ratio indicates the autophagosome-lysosome fusion efficiency. Autophagosome puncta were counted from 30–50 cells at each condition. Western blot analysis to identify the effect of starvation or chloroquine (CLQ) treated conditions in the HepG2 WT (e) and HepG2 ATP7B−/− (f) cells. Statistical differences in the studied groups were assessed as follows: * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001 and ns = not significant.