Fig. 5

AURKA modulates NKX3.1 stability via direct phosphorylation at S28, 101, and 209. A 3A-NKX3.1 shows higher expression as compared to NKX3.1 in C4-2 cells. B Quantification of NKX3.1 levels normalized to actin from three independent experiments. *P < 0.05. C 3A-NKX3.1 displays higher expression as compared to wild-type in 22Rv1 cells. D Quantification of NKX3.1 levels normalized to actin from three independent experiments in 22Rv1 cells. *P < 0.05. E CHX experiments revealed higher stability of 3A-NKX3.1 as compared to WT NKX3.1 in C4-2 cells. F Graphical representation showing mean ± SD from three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001. G CHX experiments revealed higher stability of 3A-NKX3.1 as compared to WT NKX3.1 in 22Rv1 cells. H Quantification shows mean ± SD from three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001. I 3A-NKX3.1 most efficiently ubiquitylates AURKA as compared to control C4-2 and WT NKX3.1-C4-2 cells. J 3A-NKX3.1 most efficiently ubiquitylates AURKA as compared to control 22Rv1 and WT NKX3.1-22Rv1 cells. K Ectopic overexpression of kinase-dead AURKA does not alter NKX3.1 protein levels substantially . L Quantitative analysis of three independent experiments using WT and kinase-dead (KD) AURKA reflect the protein levels of NKX3.1 in C4-2 cells, **P < 0.01 relative to control