Journal: bioRxiv

Article Title: Therapy-induced PSMA2 Sensitizes Prostate Cancer Cells to Residual Androgen and Promotes Neuroendocrine Lineage Transformation

doi: 10.1101/2025.11.25.690552

Figure Lengend Snippet: (A) Quantitative PCR graph showing fold change expression of PSMA2 mRNA normalized to GAPDH in prostate cancer cells (LNCAP or LASCPC-01 or NCI-H660) treated with enzalutamide (1uM, 48 hours) (blue bars) versus untreated controls (grey bars). For this and all subsequent graphs p-values are derived from T-test (n.s=not significant). (B) Image of a Western blot showing PSMA2 abundance in LASCPC-01 cells left untreated or treated with enzalutamide (1uM) for 48hrs. Blots were stained against PSMA2 or GAPDH as loading control. (C) Western blot image showing expression of PSMA2 in TRAMP mouse prostate tumor tissue. Twenty-five-weeks-old B6FVB TRAMP mice received regular water (Control) or water containing enzalutamide (3mg/kg) (experimental) for 6 weeks. Prostates tissues were harvested and processed for protein extraction and Western blotting. Blots were stained against PSMA2 or β-Tubulin as loading control. (D) Box plot showing PSMA2 mRNA expression in TCGA and GTEx normal (red) versus tumor (blue) prostate tissues.

Article Snippet: Blots were stained with antibodies against PSMA2 (Origene#CF505474), HSP90 (ThermoFisher Scientific # MA110372), PSA/KLK3 (Proteintech#60338-1), AR (Invitrogen # MA5-13426), and GAPDH (1:1000, DSHB-hGAPDH-2G7) or b-tubulin (1:1000, DSHB-E7).

Techniques: Real-time Polymerase Chain Reaction, Expressing, Derivative Assay, Western Blot, Staining, Control, Protein Extraction

Journal: bioRxiv

Article Title: Therapy-induced PSMA2 Sensitizes Prostate Cancer Cells to Residual Androgen and Promotes Neuroendocrine Lineage Transformation

doi: 10.1101/2025.11.25.690552

Figure Lengend Snippet: (A) Trypan blue cell viability assay showing androgen growth response of LASCPC-01 or PSMA2-KD LASCPC-01 cells. Cells were treated with R1881 (0.05 or 0.1nM) in the absence (Untreated) or presence of enzalutamide (1uM). (B) Quantitative PCR graph showing NXK3-A expression in controls LASCPC-01 cells or PSMA2-KD LASCPC-01 cells treated with increasing levels of R1881 (0.01-1nM) in the presence of enzalutamide (1uM). Expression was normalized to GAPDH mRNA. (C and D) Western blot images from anti-AR immunoprecipitation (IP) assays on lysates derived from intact or PSMA2-depleted (PSMA2-KD) LASCPC-01 (c) or NCI-H660 (d) cells. Blots were stained with antibodies against Hsp90 or AR or β-tubulin. (E) Western blot image showing Hsp90 protein abundance in intact or PSMA2-depleted LASCPC-01 before and after treatments with increasing concentration of MG132 (10, 20, 30uM). Blots were stained with antibodies against Hsp90 or GAPDH (loading control). (F and G) Western blot image showing that direct partial over-expression of PSMA2 (F) is sufficient to sensitize LNCAP cells to androgen signaling-dependent upregulation of NKX3-1 (G). (H) Western blot images of lysates derived from controls or PSMA2-overxpressing LNCAP cells. Blots were stained against PSMA2 or or HSP90 or loading controls (β-tubulin and GAPDH, respectively). (I) MTT cell viability graph showing the relative viability of unaltered or PSMA2 over-expressing LNCAP cells following enzalutamide treatment.

Article Snippet: Blots were stained with antibodies against PSMA2 (Origene#CF505474), HSP90 (ThermoFisher Scientific # MA110372), PSA/KLK3 (Proteintech#60338-1), AR (Invitrogen # MA5-13426), and GAPDH (1:1000, DSHB-hGAPDH-2G7) or b-tubulin (1:1000, DSHB-E7).

Techniques: Viability Assay, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Immunoprecipitation, Derivative Assay, Staining, Quantitative Proteomics, Concentration Assay, Control, Over Expression