Journal: Biomolecules
Article Title: Trpm8 Expression in Human and Mouse Castration Resistant Prostate Adenocarcinoma Paves the Way for the Preclinical Development of TRPM8-Based Targeted Therapies
doi: 10.3390/biom12020193
Figure Lengend Snippet: Cooperative effect of the Trpm8 agonist D−3263 with Enzalutamide or Docetaxel in TRAMP PCa cell lines. ( A , B ) End−point PCR ( A ) and Western blotting ( B ) analyses showing the expression of Trpm8 in TRAMP C1 and C2 cell lines compared to normal mouse prostate AP, DLP and VP lobes. Gapdh was used as loading control. Experiments were performed in quadruplicate. ( C ) Western blotting analysis showing molecular signature of apoptotic cell death (Caspase−3 and Parp cleavage). TRAMP C1 and TRAMP C2 cells were treated with D−3263 (1 μM), Docetaxel (5 nM), Enzalutamide (1 μM), D-3263 + Docetaxel, or D-3263 + Enzalutamide for 24 or 72 h. Untreated cells were used as control. Gapdh was used as loading control. Experiments were performed in triplicates. * Protein extracts from cells treated with Staurosporine (1 μM, 6 h) were used as positive control.
Article Snippet: Then, the Trpm8 response to D-3263 was evaluated by measuring cytosolic calcium accumulation through quantitative Fluo-4 imaging.
Techniques: Polymerase Chain Reaction, Western Blot, Expressing, Positive Control