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. 2022 Apr 9;52:102312. doi: 10.1016/j.redox.2022.102312

Fig. 7.

Fig. 7

HIF-1α to regulate ELAVL1 cytoplasmic translocation and the mRNA stability of SLC7A11 in hypoxic environment by PMAN.a MGC-803 cells were cultured in hypoxia, RT-qPCR analyzes the expression level of SLC7A11 mRNA at indicated timepoints. Mean ± SD is shown. b MGC-803 cells were cultured in hypoxia, western blotting analyzes the expression level of SLC7A11 at indicated timepoints. c RT-qPCR detects the expression level of SLC7A11 mRNA in siHIF-1α-transfected MGC-803 cells. Mean ± SD is shown. Statistical analysis was conducted using Student's t-test. d Correlations between HIF-1α mRNA levels and SLC7A11 mRNA levels in GC. R-values and P-values were used via Pearson's correlation analysis. e The expression of SLC7A11 in siHIF-1⍺ or siNC transfected MGC-803 cells, PMAN-overexpression MGC-803 cells co-transfected with siHIF-1⍺ or siNC and transfected siHIF-1⍺ or siNC in PMAN-overexpression MGC-803 cells co-transfected with siELAVL1 or siNC after 24 h under hypoxia. fSLC7A11 mRNA decay line chart in MGC-803 cells transfected with siHIF-1⍺ or siNC during hypoxia. Mean ± SD is shown. Statistical analysis was conducted using one-way ANOVA. gSLC7A11 mRNA decay line chart of PMAN-overexpression MGC-803 cells co-transfected with siHIF-1⍺ or siNC during hypoxia. Mean ± SD is shown. Statistical analysis was conducted using one-way ANOVA. h Protein nuclear-cytoplasmic fractionation assay showed the expression level of ELAVL1 in the subcellular fractions of PMAN-NC MGC-803 cells under hypoxia or normoxia and PMAN-NC MGC-803 cells transfected with siHIF-1⍺ or siNC under hypoxia by western blotting. Similarly, the expression level of ELAVL1 in the subcellular fractions of PMAN-overexpression MGC-803 cells under hypoxia or normoxia and PMAN-overexpression MGC-803 cells transfected with siHIF-1⍺ or siNC under hypoxia. Protein quantification on ImageJ and Histone H3 was treated as a nuclear control while β-tubulin was a cytoplasmic control. Mean ± SD is shown. Statistical analysis was conducted using one-way ANOVA. i IF showed ELAVL1 cytoplasmic fluorescence intensity (green) in transfected siHIF-1⍺ or siNC MGC-803 cells and PMAN-overexpression or PMAN-NC MGC-803 cells transfected with siHIF-1⍺. The nucleus was stained with DAPI. Scale bar, 20 μm. 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.)