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. 2023 Jul 14;9:247. doi: 10.1038/s41420-023-01506-x

Fig. 2. FKBP5 inhibition protected β cells against proinflammatory cytokines-induced apoptosis via autophagy regulation.

Fig. 2

A NIT-1 cells were transfected with siFkbp5 or control siRNAs and then treated with cytokines, mRNA expression of Nkx6.1 and Mafa were evaluated by qRT-PCR. Experiments were repeated for 3 times. B, C mRNA expression of Nkx6.1 and Mafa in NIT-1 cells (B) and primary human islets (C) treated with proinflammatory cytokines (Cytokines), or cytokines together with SAFit2 (Cytokines + SAFit2). Experiments were repeated for 3 times. D, E Western blot with LC3, p62 and Bax in the cytokines, or cytokines+ SAFit2-treated NIT-1 cells (D). The signal intensity was quantified by Image J with β-actin as loading control. Experiments were repeated for 3 times (E). F, G Annexin-V/7-AAD staining and flow cytometry analysis in NIT-1 cells treated with cytokines, cytokines + SAFit2, or Cytokines + SAFit2 + CQ) (F). Quantification of the apoptotic cell rate (PE+7AAD+/PE+7AAD-). Experiments were repeated for 3 times (G). H, I Autophagy level measurement by flow cytometry in NIT-1 cells treated with Cytokines, or Cytokines + SAFit2. CQ was used to allow the accumulation of autophagic vacuoles. FITC-A intensity represents the autophagy level, Experiments were repeated for 3 times.. Student’s t-test. Mean ± SEM, *P < 0.05, **P < 0.01, ***P < 0.001.