Fig. 1: O-GlcNAcylation increases ferroptosis sensitivity of HCC cells.

A and B Cell viability was measured by an MTT assay in SMMC-7721 cells treated with RSL3 at the indicated concentration (A) and for the indicated time (B). C Global O-GlcNAcylation was measured by WB after PUGNAc treatment (25 µM). D The fluorescence of C11-BODIPY was measured to detect lipid ROS in Bel-7402 and SMMC-7721 cells treated as indicated. E MDA levels were measured in Bel-7402 and SMMC-7721 cells treated as indicated. F Global O-GlcNAcylation was detected by WB in cells treated with PUGNAc and RSL3 separately or simultaneously. G–I Ferroptotic events were evaluated in cells treated with PUGNAc and RSL3 separately or simultaneously: cell death (G), lipid ROS production (H), and MDA production (I). J Global O-GlcNAcylation and OGT expression were detected by WB in Bel-7402 and SMMC-7721 cells overexpressing OGT. Cells were subsequently treated with RSL3 as indicated. K–M Ferroptotic events were evaluated in cells overexpressing OGT and treated with RSL3 as indicated: cell death (K), lipid ROS production (L), and MDA production (M).