Fig. 4: P4HA3 confer ferroptosis resistance through promoting SLC7A11 expression.

A RNA-seq was performed using SiHa-PR cell lines with or without overexpression of P4HA3. Heatmap showing the top 10 differentially expressed genes between control and P4HA3-overexpression cells (expect for P4HA3). Three samples were used in each group. Colors correspond to the expression level indicated by the log2-transformed scale bar below the matrix. B RT-qPCR and western blot analysis of SLC7A11 expression in P4HA3-overexpressing HeLa PR and SiHa PR cells. β-actin was the internal control for RT-qPCR. (n = 3). C RT-qPCR and western blot analysis of SLC7A11 expression in P4HA3-silenced HeLa LNM2 and SiHa LNM2 cells. β-actin was the internal control for RT-qPCR. (n = 3). D Correlation analysis between P4HA3 and SLC7A11 using a cohort of 60 cervical cancer tissues which were obtained from patients who underwent gynecological surgery at the First Affiliated Hospital of Sun Yat-sen University. E, F The mRNA and protein expression of SLC7A11 were detected by RT-qPCR and western blot. G, H Flow cytometry of C11-BODIPY staining in the indicated group. (n = 3). I, J MDA and GSH levels in shNC+Vector, shP4HA3+Vector, and shP4HA3 + SLC7A11 CCa cells. (n = 3). K TEM images showing mitochondrial morphology in indicated cells. L, M CCa cells were implanted subcutaneously into the footpad region of female nude. Six weeks later, popliteal LNs were dissected for images and quantification of LN volumes in indicated groups (n = 8). N, O IF staining of pan-cytokeratin (CK) and quantification of metastatic areas in popliteal LN. P, Q Representative images and quantification of 4-HNE staining in primary tumors. Error bars represent the means ± SD. ns no significance; *p < 0.05; **p < 0.01; ***p < 0.001; and ****p < 0.0001.