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. 2022 Apr 6;298(5):101904. doi: 10.1016/j.jbc.2022.101904

Figure 8.

Figure 8

Carbamazepine (Carb) prevents mutant KATPchannels from autophagy-dependent protein degradation.A, top, representative immunoblots for LC3BI, LC3BII, and p-AMPKα1/2 in INS-1 cells expressing WT (HA-SUR1/Kir6.2) or mutant (HA-SUR1/Kir6.2-A28V) KATP channels, following 10 μM Carb treatment for 16 h. Bottom, the bar graphs show quantification of the LC3BII/LC3BI ratio and p-AMPKα1/2. B, top, representative immunoblots for p62 in INS-1 cells expressing WT (HA-SUR1/Kir6.2) or mutant (HA-SUR1/Kir6.2-A28V) KATP channels, following 10 μM Carb treatment for 16 h. Bottom, the bar graphs show quantification of p62. F, top-left, representative immunoblots show the levels of pS6 and SUR1 in INS-1 cells expressing WT or mutant KATP channels, after treatment with vehicle, 10 μM Carb, 25 μM rapamycin, or 25 μM rapamycin + 10 μM Carb for 16 h. Top-right, the bar graph shows the levels of pS6. Bottom, quantification of the upper (mature) and lower bands of SUR1 is shown in the left and right plots, respectively. Data are presented as the mean ± SEM; n = 3 per group; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗∗p < 0.001, compared with the WT-vehicle group; differences were evaluated using two-way ANOVA and Dunnett's post hoc test. AMPK, AMP-activated protein kinase; HA, hemagglutinin; KATP, ATP-sensitive potassium channel; SUR1, sulfonylurea receptor 1.