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Fig. 4 | Diabetology & Metabolic Syndrome

Fig. 4

From: Targeting ferroptosis with miR-144-3p to attenuate pancreatic β cells dysfunction via regulating USP22/SIRT1 in type 2 diabetes

Fig. 4

USP22 suppressed HG-induced ferroptosis in pancreatic β cells by stabilizing SIRT1 expression. A Co-IP was performed in cells transfecting with Flag-tagged USP22 and Myc-tagged GPX4 or HA-tagged SIRT1. Cell lysates were immunoprecipitated with anti-Flag antibodies. B In vitro ubiquitination assay was performed in cells transfecting with HA-tagged SIRT1 alone or together with Flag-tagged USP22; After transfection 48 h, cells were treated with MG132 for 6 h. Cell lysates were subjected to immunoprecipitated with anti-HA antibodies. C Cells were transfected with USP22 OE and harvested at 0, 2, 4, and 8 h after treatment with CHX to detect the expression of SIRT1 by western blot. RT-qPCR detected the transfection efficiency of D SIRT1 overexpression vector (SIRT1 OE) and E USP22 short hairpin RNA (sh-USP22#1 and sh-USP22#2). F The levels of SIRT1, USP22, and ferroptosis-related proteins (GPX4 and TFR1) were evaluated by western blot (***P < 0.005 vs. vector; ###P < 0.005 vs. SIRT1 OE)

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Diabetology & Metabolic Syndrome

ISSN: 1758-5996

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