Fig. 4

ZMYND11 antagonizes oncogenic function of HNRNPA1 in prostate cancer dependent on its MYND ___domain. a Knockdown efficiency of shRNAs against HNRNPA1 determined by western blot analysis. b Colony-forming units of 22Rv1 cells stably expressing control or shRNAs against HNRNPA1. c Proliferation capacity of 22Rv1 cells measured at the indicated times. Absorbance at 450 nm; mean ± SD of three independent experiments. Representative images and quantification of relative migration (d) and invasion (e) for the cells stably expressing the indicated shRNAs. Error bars, ±SD of triplicate experiments. f HNRNPA1 expression levels are significantly upregulated in human metastatic prostate tumors. P values determined by the Wilcoxon and Kruskal-Wallis tests, respectively. g HNRNPA1 upregulation correlates with higher Gleason grade cancer. P values assessed by the Kruskal-Wallis test. h Immunohistochemistry on the expression of HNRNPA1 and evaluation of HNRNPA1 staining intensity in a paraffinized prostate tissue microarray consisting of 163 samples. Original magnification, 100×; insets, 400×; Scale bar, 100 μm. i Immunostaining for HNRNPA1 was performed on another cohort of prostate specimens. Representative images and quantification of HNRNPA1 protein staining are shown. Original magnification, 400×; Scale bar, 100 μm. j Kaplan-Meier analysis of overall survival in the Tongji cohort of patients with prostate cancer. Patients were stratified into two groups according to the state of HNRNPA1 expression, higher (top 50%; n = 35) or lower (bottom 50%; n = 35). k Kaplan-Meier analysis showed that HNRNPA1 upregulation is significantly associated with elevated risk of biochemical recurrence or metastasis in prostate cancer subsets within the TCGA database. l High expression levels of HNRNPA1 indicates predictive values for recurrence-free survival in patient group with Gleason sum score 7 prostate cancer (intermediate-risk). Representative images and quantification of colony formation (m), cell proliferation (n), migration (o), and invasion (p) for 22Rv1 cells transfected with empty vector, HNRNPA1 alone, or together with the indicated full-length ZMYND11 or MYND-___domain deletion mutant. Scale bar, 400 μm. q Kaplan-Meier analysis of biochemical recurrence or metastasis-free survival in the TCGA cohort of prostate cancer patients stratified into two groups with ZMYND11^low/HNRNPA1^high and ZMYND11^high/HNRNPA1^low expression, respectively. Error bars, ±SD (b–e, m–p). ns not significant, *p < 0.05, **p < 0.01, ***p < 0.001, two-tailed Student’s t tests