Fig 2: CPSF4 gene set enrichment study from prostate cancer patients’ TCGA-PRAD RNA-seq data. (A) Heatmap analysis showing the top 50 associated genes for each of the two groups of most up- and down-regulated genes, a total of 20,051 genes were analyzed. (B) Gene ontology (GO) enrichment analysis showing the biological, cellular, and molecular processes that are most positively and negatively associated, respectively. FDR values and Pearson correlation coefficients were used to rank the data. Using the Benjamini–Hochberg approach, the FDR is computed. (C) Heatmap linking the most up- and down-regulated proteins to the RNA expression of CPSF4 from patients in TCGA database.

Fig 3: Dysregulation of cell cycle progression and signaling results from CPSF4 knockdown. (A) Analysis of the cell cycle and apoptosis using three replicates of flow cytometry on DU-145 control and siCPSF4-treated cells. (B) Analysis of the cell cycle and apoptosis in the siCPSF4-treated PC3 group versus the untreated control group. (C) Pictures of PC3 and DU-145 cell lines treated with siCPSF4 and scramble siRNA (negative control) from a colony formation test. (D) A graphic representation of a quantified analysis of a colony formation experiment (Astericks *** indicates p < 0.001 and **** indicates p < 0.0001). Dark column represents control trial and lighter/checkered column shows knockdown trial.

Fig 4: Prostate cancer cells are less able to move and invade when CPSF4 is inhibited. (A) Data of the invasion assay for the PC3 and DU-145 cell lines treated with siCPSF4, and scramble RNA (negative control) are shown. The scale bar indicates 400 μm, and the data were obtained from 3 repetitions. (B) Quantification of the invasive cell population from the PC3 and DU-145 cells invasion assay. (C) Images of the migration assay findings for the PC3 and DU-145 cell lines from three duplicate groups, including both control and CPSF4 knockdown treatment groups. (D) A graphic depiction of PC3 and DU-145 cell migration numbers in control and siCPSF4-treated cells. Blue graphs depict DU-145 cells and red graphs depict PC3 cells. (Asterisk **** indicates p < 0.0001). Scale bar in the figure represents 400 μm.

Fig 5: Analysis of CPSF4 using a PCa cell line Western blot. Blue graphs depict DU-145 cells and red graphs depict PC3 cells. (A) Western blot of the CPSF4 expression in the HEK-293, RWP-1, DU-145, PC3, and PC3-ERG cell lines, which are Pca cell lines. (B) After 48 h, the effectiveness of CPFS4 knockdown utilizing CPSF4 siRNA #1, #2, and scrambled siRNA as a negative control. (C) Western blotting study of the expression of the proteins E-Cadherin, Vimentin, c-Myc, and β-catenin in CPSF4 knockdown PC3 and DU-145 cells in comparison to control cells. GAPDH was employed as an internal quality control measure. (D) Graphical representation of c-Myc, β-catenin, ECAD, and Vimentin expression in siCPSF4 and control trials for DU-145 and PC3 cell lines (Asterisk ** p< 0.01 and *** indicates p < 0.001).