Anti blm

PC3 cells were seeded in a six-well plate and incubated for 48 h. Cells were then fixed using 3.7% paraformaldehyde and permeabilized with 0.25% Triton X-100. Subsequently, blocking was carried out with 3% Bovine Serum Albumin (BSA). Cells were incubated with a primary antibody on a shaking table at 4 °C overnight. This was followed by incubation with a Cy3-conjugated secondary antibody at room temperature for 2 h. The primary antibody utilized was anti-BLM (dilution ratio of 1:150; Bioss, China), and the secondary antibody was fluorescent Cy3-conjugated goat anti-rabbit antibodies (dilution ratio of 1:600; Invitrogen, Carlsbad, CA, USA). Fluorescence imaging was conducted using a fluorescence microscope (Nikon, Japan).

Cisplatin (HY-17394), Z-VAD-FMK (HY-16658), doxorubicin (HY-15142), and staurosporine (62996-74-1) were purchased from MCE (Middlesex, NJ, USA). ML216 was purchased from MCE (HY-12342) and Selleck (S0469, Houston, TX, USA). Hydroxyurea (V900323) and etoposide (E1383) were purchased from Sigma (St. Louis, MO, USA). The antibodies used included anti-DBC1 (Bethyl Labotatories, Montgomery, TX, USA, A300-434A), anti-V5 (Invitrogen, Waltham, MA, USA, R960-25), anti-Flag (Sigma, F7425), anti-BLM (Bethyl Labotatories, A300-110A), anti-GAPDH (Millipore, St. Louis, MO, USA, MAB374), anti-p21 (BD Biosciences, San Jose, CA, USA, 556430), anti-β Tubulin (Santa Cruz, Dallas, TX, USA, SC-166729), anti-p53 (Santa Cruz, SC-126), anti-Bcl-2 (Santa Cruz, SC-7382), anti-p16 (Santa Cruz, SC-56330), anti-phospho-H2A.X (Millipore, 07-164-25UG), anti-β-actin (Absin, Shanghai, China, ABS830031), anti-PARP1 (CST, Danvers, MA, USA, 9542S), anti-cleaved PARP1 (CST, 5625S), anti-BLM (Bioss, Beijing, China, bs-12872R) and anti-p21 (Bioss, bs-0741R). Secondary antibodies were horseradish peroxidase-coupled sheep anti-mouse IgG (GE Healthcare, Chicago, IL, USA, NA931 or ZSGB-BIO, Beijing, China, ZB-2301), donkey anti-rabbit IgG (GE Healthcare, NA934), and horseradish peroxidase-coupled sheep anti-mouse IgG (ZSGB-BIO, ZB-2305).