Fig. 3: Overexpression of HIF-1α improves ferroptosis in inflammatory colon epithelial cells.
From: HIF-1α alleviates ferroptosis in ulcerative colitis by regulation of GPX4
A Immunofluorescence detection of HIF-1α and FTH in NCM460 cells. Nuclei were stained with DAPI in blue, HIF-1α localization was indicated in green, and FTH localization was indicated in red (Scale: 25 μm). B, C Western blot and RT-qPCR analysis of HIF-1α in wild type (WT), empty control (Vector), and overexpression group (HIF1A-OE), with β-actin as internal reference. D Western blot detection of tight junction proteins (Occludin and ZO-1), zonulin, and HIF-1α expression in intestinal epithelial cells (IECs) of different groups. E The mRNA levels of HIF-1α, IL-6, and IL-1β in cells from each group with and without LPS intervention. F, G Evaluation of intestinal epithelial cell integrity by FITC-dextran permeability assay and transepithelial electrical resistance (TEER). H Flow cytometry detection of C11-BODIPY581/591 levels in IECs with or without LPS intervention. I MDA levels in human IECs using MDA kit. J Iron levels of human IECs in each group were determined by iron assay kit. K Western blot detection of iron storage proteins (FTL and FTH) and HIF-1α expression in IECs. L Flow cytometric detection of PI-positive cells to analysis necrotic cell death in IECs. M Transmission electron microscopy of human IECs under different intervention conditions (Scale: 1 μm, 5 μm). N The protein expression levels of ACSL4, LPCAT3, SLC7A11, GPX4, TXNRD1, and HIF-1α in human IECs under different treatments as assessed by Western blot. ns P > 0.05, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.
