Fig. 3

PACT regulates PCa cell cycle progression. (A, B) DAVID gene ontology (GO) enrichment analyses of the pathways downregulated (A) and upregulated (B) with PACT KO; (i) GO Biological processes, and (ii) KEGG pathways. (C) Gene Set Enrichment Analysis (GSEA) of the RNA-seq (i) biological processes; and (ii–iii) representative enrichment score plots. The y-axis and the green line show the enrichment score for each gene, illustrated as a vertical line plotted in rank order of the most gene abundance (red, left) to the least gene abundance (blue, right) within the indicated samples (as log₂FC/comparison); the black vertical lines correspond to member genes from the set. NES = normalized enrichment score, FDR = false discovery rate. Pathways ranked by the p-value result of a Fisher’s exact test. (D–E) Flow cytometry cell cycle analysis (D) and apoptosis analysis (E) of LNCaP cells ± PACT; (i) LNCaP cells transfected with si-NC or si-PACT (20 nM) for 72 h, and (ii) LNCaP parental and LNCaP PACT KO cells assayed 72 h post-plating. n = 3; data analysis was with an unpaired two-tailed student’s t-test; with significance denoted as: *p < 0.05, **p < 0.005 relative to si-NC or parental cells. See Supplementary Fig. S3C (i-ii) for RT-qPCR validation of si-PACT knockdown.