Fig 1: NEK6-KO increases ROS levels and its expression modulates antioxidant defenses in DU-145 cells. (A) DU-145 WT and NEK6-KO’s ROS levels were measured by DHE staining in flow cytometry. NEK6-KO showed an increase in ROS levels when compared with WT cells. (B) Correlation of the gene expression in prostate adenocarcinoma (PRAD) using the GEPIA platform. Spearman’s coefficient was used to measure the intensity of the correlation between genes. (C) mRNA levels of SOD1, SOD2, and PRDX3 were analyzed through RT-qPCR. A significant reduction in mRNA of SOD2 was observed. (D) The expression of antioxidant proteins (SOD1, SOD2, and PRDX3) was evaluated by Western blotting. Target deletion of the NEK6 gene reduces the expression of SOD1, SOD2, and PRDX3. Additionally, high levels of JNK phosphorylation were observed in NEK6-KO cells. (E) Quantification of antioxidant protein expression in WT and NEK6-KO cells. (F) NEK6 overexpression in DU-145 WT cells upregulated the SOD1, SOD2, and PRDX3 protein expression. (G) Quantification of antioxidant protein expression in FLAG-NEK6 expressing cells. * p < 0.05, ** p < 0.01 and *** p < 0.001.