Fig. 7: KLF5 retains oncogenic function and activates ERBB2 in an enzalutamide-resistant CRPC cell line model of AR-V activity.
From: Opposing transcriptional programs of KLF5 and AR emerge during therapy for advanced prostate cancer
a KLF5 protein measured by western blot using R1-D567 cells infected with control shRNA lentivirus (shC) or two independent KLF5-targeted shRNA lentiviruses (shK1 and shK2). ERK2 is a loading control. b R1-D567 cells as in a analyzed by 3D soft agar colony formation assays. n = 14, mean ± 95% CI from four independent experiments, two performed in biological triplicate and two performed in biological quadruplicate. P-values are unadjusted from 2-sided, 2-tailed t-tests. c R1-D567 cells transfected with KLF5-targeted siRNAs (siK1 and siK2) or control siRNA (siC) analyzed by chemotactic migration assays. n = 6, mean ±95% CI, two independent experiments in biological triplicate. P-values are unadjusted from 2-sided, 2-tailed t-tests. d Heatmaps of AR variant (AR-V) and KLF5 ChIP-seq signals ±3 kb around KLF5/AR common, KLF5 only, or AR only peaks in R1-D567 cells cultured in androgen-deplete medium. e Heatmap of RNA-seq gene expression data for a set of 79 genes located ±50 kb from a KLF5/AR common peak and differentially expressed in R1-D567 cells based on comparisons between shRNA control (shCTRL) vs. shKLF5 conditions and/or DHT vs. vehicle conditions. The heatmap was generated by unsupervised clustering, revealing two main clusters. f GSEA testing enrichment of the signature ERBB2_UP.V1_UP in R1-D567 gene expression data reflecting active KLF5 (differential expression in shCTRL vs. shKLF5).
