Fig. 4.

PMAN affected mRNA stability of SLC7A11 to enhance the expression of SLC7A11.a The expression level of ferroptosis-related proteins in the PMAN knockdown AGS cells and the PMAN overexpression MGC-803 cells. b Immunohistochemistry was used to analyze SLC7A11 expressions in 3 pairs GC peritoneal metastasis samples. Scale bar, 20 μm cSLC7A11 mRNA expressions in 30 pairs GC samples. Mean ± SD is shown. Statistical analysis was conducted using Student's t-test. dSLC7A11 mRNA expressions in 3 pairs GC peritoneal metastasis samples. Mean ± SD is shown. Statistical analysis was conducted using one-way ANOVA. e SLC7A11 expression levels in 6 pairs GC samples and 3 pairs GC peritoneal metastasis samples. f Correlations between SLC7A11 mRNA levels and PMAN levels in GC. R-values and P-values were used via Pearson's correlation analysis. g The location of PMAN (red) was detected by FISH assay in AGS cells. The nucleus was stained with DAPI. Scale bar, 20 μm h RNA nuclear-cytoplasmic fractionation assay showed the expression level of PMAN in the subcellular fractions of AGS cells by RT-qPCR. U6 was treated as a nuclear control while β-actin was a cytoplasmic control. Mean ± SD is shown. i SLC7A11 was detected by western blotting in PMAN-overexpression MGC-803 cells after treated with CHX (10 μM). j-kSLC7A11 mRNA decay line chart of PMAN-overexpression MGC-803 cells (j) and PMAN knockdown AGS cells (k) after treated with actinomycin D (5 μg/ml). Mean ± SD is shown. Statistical analysis was conducted using one-way ANOVA. Ns = nonsignificant (p > 0.05), *p < 0.05, **p < 0.01. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)