Fig. 1: Pro-inflammatory cytokines stimulate the AMPK-ULK-1 axis while inhibiting mTORC1 in β-cells | Cell Death & Disease

Fig. 1: Pro-inflammatory cytokines stimulate the AMPK-ULK-1 axis while inhibiting mTORC1 in β-cells

From: Dysfunctional autophagy following exposure to pro-inflammatory cytokines contributes to pancreatic β-cell apoptosis

Fig. 1

a–d Prevalence of apoptosis was evaluated by HO-PI staining in INS-1E cells a, c or primary rat islets b, d treated or not (ctrl) for 16 h with IL-1β + IFNγ (cyt), alone or in combination with chloroquine (CQ; 10 µM), Bafilomycin A1 (Baf; 100 nM), 3-Methyladenine (3-MA, 5mM), rapamycin (rap; 100 nM), or torin1 (1–100 nM, as indicated). Data are mean ± SEM of 4–6 independent experiments. *P < 0.05;**P < 0.01; ***P < 0.001 vs. ctrl. #P < 0.05; ##P < 0.01 vs. IL-1β + IFNγ as determined by two-way ANOVA with Dunnet’s correction for multiple comparisons. e INS-1E cells infected with the Ad-DN-ULK-1 were treated or not (ctrl) with IL-1β + IFNγ (cyt) for 15 h. Prevalence of apoptosis was evaluated by HO-PI staining. Data are mean ± SEM of four independent experiments. **P < 0.01; ***P < 0.001 vs. respective ctrl condition (white bars). #P < 0.05; ##P < 0.01 vs. respective Ad-GFP condition as determined by two-way ANOVA with post-hoc t-test with Sidak’s correction for multiple comparisons. f Upper panel: Western blot analysis of the ATG5–12 complex and tubulin in INS-1E cells transfected or not (NT) with a control siRNA (siCtrl) or two siRNA targeting ATG5 (siATG51 and 2). Lower panel: prevalence of apoptosis in transfected INS-1E cells treated or not (ctrl) with cytokines for 24 h (cyt). Data are mean ± SEM of three independent experiments.*P < 0.05 vs. respective ctrl (white bars); #P < 0.05, ##P < 0.01 vs. respective siCtrl condition as determined by two-way ANOVA with post-hoc t-test with Sidak’s correction for multiple comparisons. g Time-course Western blot analyses of P-AMPK, P-ULK-1, P-Raptor, LC3-I and II and tubulin in INS-1E cells treated with IL-1β + IFNγ. Data are representative of four independent experiments. h Western blot analysis of P-AMPK, P-Raptor, P-ULK-1, LC3-I and II and tubulin in primary rat islets exposed for 24 h to IL-1β + IFN-γ (cyt). Data are representative of four separate islet preparations

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