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. 2012 Jun 11;122(7):2469–2481. doi: 10.1172/JCI62044

Figure 5. PXN modulates ERK-mediated transcription independent of androgens.

Figure 5

(A) PXN is required for EGF-induced cyclin D1 promoter activity. Nsp/PXN-specific siRNA-treated PC3 cells were transiently transfected with cyclin D1 promoter-luciferase plus cytomegalovirus–β-gal plasmid, followed by EGF stimulation (20 ng/ml; 24 hours). PXN-knockdown cells were transfected with plasmids expressing WT PXN or PXN lacking the MAPK-targeted serines (S→A). Cyclin D1 promoter-luciferase activity was normalized to β-gal expression and data represented as fold increase with respect to medium (mean ± SEM, n = 3). *P ≤ 0.001. (B) Nuclear localization of ERK is independent of PS-PXN. Immunofluorescence of ERK (red), PXN (green), p-ERK (green), or PS-PXN (green) in PC3 cells treated with Nsp siRNA or PXN siRNA, followed by medium or EGF (20 ng/ml; 30 minutes) stimulation. PXN-knockdown cells were transfected with plasmids expressing WT PXN or PXN lacking the MAPK-targeted serine (S→A). Adjacent Hoechst staining represents the nucleus (n = 5 experiments, all with identical results). Original magnification, ×40.