Fig. 3.

The overexpression of PMAN inhibits ferroptosis and knockdown of PMAN promotes ferroptosis in GC cells.a Cell morphology of AGS cells under an inverted microscope after treated with Erastin (10 μM) or RSL3 (2 μM) at indicated timepoints. b Transmission electron microscopy (TEM) was performed to evaluate the change in ultrastructural features such as mitochondria of AGS cells under Erastin or RSL3 treatment after 12 h. Scale bar, 5000 nm c RT-qPCR analysis was used to detect the expression of PMAN in MGC-803. Mean ± SD is shown. Statistical analysis was conducted using Student's t-test. d CCK-8 assay to analyze the effect of overexpression-PMAN on cell proliferation in MGC-803. e Level of total iron f level of ferrous iron g level of lipid ROS h the ratio of GSSG/GSH i level of MDA were detected in MGC-803 cells stably overexpressing PMAN under Erastin (10 μM) or RSL3 (2 μM) treatment after 12 h. Mean ± SD is shown. Statistical analysis was conducted using Student's t-test. j Transmission electron microscopy was performed to evaluate the change of mitochondrial ultrastructure in MGC-803 cells stably overexpressing PMAN after treated with Erastin (10 μM) or RSL3 (2 μM). Scale bar, 5000 nm k RT-qPCR analysis was used to detect the expression of PMAN in AGS cells. Mean ± SD is shown. Statistical analysis was conducted using Student's t-test. l CCK-8 assay to analyze the effect of knockdown-PMAN on cell proliferation in AGS cells. m Level of total iron n level of ferrous iron o level of lipid ROS p the ratio of GSSG/GSH q level of MDA were detected in AGS cells stably knockdown PMAN under Erastin (10 μM) or RSL3 (2 μM) treatment after 12 h. Mean ± SD is shown. Statistical analysis was conducted using Student's t-test. r Transmission electron microscopy was performed to evaluate the change of mitochondrial ultrastructure in AGS cells stably knockdown PMAN after treated with Erastin (10 μM) or RSL3 (2 μM). Scale bar, 5000 nm. Ns = nonsignificant (p > 0.05), *p < 0.05, **p < 0.01.