Fig. 5

ZMYND11 regulates HNRNPA1-mediated alternative splicing of PKM and counteracts PKM2-induced aggressive cellular phenotype in prostate cancer. RNA (a, c) or protein (b, d) was extracted from 22Rv1 cells either transfected with full-length ZMYND11 or the indicated mutant (a, b) or stably expressing the ZMYND11-targeting shRNAs (c, d) followed by the examination of PKM isoforms both at the mRNA (a, c) and protein levels (b, d). 22Rv1 cells transfected with HNRNPA1 alone or together with either full-length ZMYND11 or the MYND-___domain-deletion mutant were assayed for PKM splicing at the mRNA (e) and protein levels (f). Representative images and quantification of colony forming (g), cell proliferation (h), migration (i), and invasion (j) in 22Rv1 cells co-transfected with PKM2 alone or together with either the full-length ZMYND11 or the MYND-___domain-deletion mutant. Mean ± SD of triplicate experiments. *p < 0.05, **p < 0.01, ***p < 0.001, Student’s t test. Scale bar, 400 μm. Relative glucose uptake and lactate production in 22Rv1 cells overexpressing ZMYND11 or its MYND-___domain-deletion mutant (k) and stably expressing ZMYND11 shRNA (l) compared to control cells. Error bars, ±SD (k–l), n = 3 biological replicates. *p < 0.05, **p < 0.01, ***p < 0.001, Student’s t tests. m Visualization of PKM splicing using LeafViz examined by RNA sequencing in 22Rv1 cells stably expressing either ZMYND11-targeting or control shRNAs. Statistic assessment via two-tailed Student’s t test. n Visualization of PKM isoform expression in 134 patients of prostate tumors and matched normal tissues in CPGEA cohort by RNA-seq profiling [PMID: 32238934]. Differential expression analysis of PKM1 (o) or PKM2 (p) in this patient cohort of 134 tumor-normal paired prostate specimens. The data were examined by the Wilcoxcon test compares two paired groups