primary antibody against prostate specific antigen  (Boster Bio)

Journal: Prostate International

Article Title: A novel biguanide derivative, IM176, induces prostate cancer cell death by modulating the AMPK-mTOR and androgen receptor signaling pathways

doi: 10.1016/j.prnil.2022.11.003

Figure Lengend Snippet: Effects of IM176, phenformin, and metformin on the inhibition of AR, AR-V7, and prostate-specific antigen (PSA) expression. (a) LNCaP and 22Rv1 cells were treated with IC 50 concentrations of IM176 (20 μmol/L and 40 μmol/L, respectively), phenformin (40 μmol/L and 400 μmol/L, respectively), and metformin (4 mmol/L and 10 mmol/L, respectively) for 72 hours, and relative mRNA levels of AR and AR-V7 were quantified by real-time PCR. Each point represents the mean ± standard deviation of triplicate determinations. ∗p < 0.05 compared with untreated control. (b) LNCaP and 22Rv1 cells were treated with the indicated concentrations of IM176, phenformin, and metformin for 72 hours. Western blot analysis was performed using antibodies to AR, AR-V7, and PSA, with GAPDH used as a loading control. (c) LNCaP and 22Rv1 cells were treated for 72 hours with the IC 50 concentrations of IM176, phenformin, and metformin, as shown in (a), above. Samples were stained with antibody to AR (red) and the nuclear stain DAPI (blue). Images were captured on a fluorescence microscope. Scale bar, 50 μM. AR, androgen receptor; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; mRNA, messenger RNA; PCR, polymerase chain reaction.

Article Snippet: Primary antibodies against phosphor-AMPK threonine 172 (pAMPK), AMPK, phospho-mTOR serine 2448 (pmTOR), mTOR, phospho-p70S6 kinase 1 threonine 389 (pp70S6K1), p70S6K1, phospho-S6 serine 235/236 (pS6), S6 caspase-3, and poly (ADP-ribose) polymerase were obtained from Cell Signaling Technology (Danvers, MA, USA); primary antibodies against AR, prostate-specific antigen (PSA), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) were from Santa Cruz Biotechnology (Dallas, TX, USA), and primary antibody against AR-V7 was from Precision Antibody (Columbia, MD, USA).

Techniques: Inhibition, Expressing, Real-time Polymerase Chain Reaction, Standard Deviation, Control, Western Blot, Staining, Fluorescence, Microscopy, Polymerase Chain Reaction