Poly l lysine coated glass

We carried out the TUNEL assay using the DEADEND Fluorometric TUNEL System (Promega, Madison, WI, USA) to evaluate effect of TRIM36 knockdown for apoptosis in PC cells. Prostate cancer cells (3.0 × 10⁴ per well) were seeded in 6‐well culture plates and incubated for 24 hours. Cells were transfected with siRNAs as described elsewhere. After incubating for 24 hours, control cells were treated with DTX (1 nmol/L) and replated to poly‐l‐lysine coated glass (Matsunami Glass Industries, Osaka, Japan) inside a 24‐well culture plate. Seventy‐two hours after transfection, cells were treated with TUNEL staining according to the manufacturer's protocol. Nuclear staining was carried out using DAPI. Signals were captured using a digital microscope (Fluoview FV10i; Olympus, Tokyo, Japan). Percentages of apoptotic cells were evaluated in 3 randomly selected fields (×100), and data are presented as mean value ± SD.

Terminal deoxynucleotidyl transferase‐mediated dUTP nick end labeling (TUNEL) assay was performed using the DEADEND Fuorometric TUNEL System (Promega, Madison, WI, USA). Cells (1.0 × 10⁵ per well) were seeded in 6‐well culture plates and incubated for 24 h. Cells were transfected with siRNAs as described, and were re‐plated to Poly‐l‐Lysine coated glass (Matsunami Glass Ind., Osaka, Japan) inside a 24‐well culture plate. Forty‐eight hours after transfection, cells were then treated with TUNEL staining according to the manufacturer's protocol. The slides were treated with 4′6′‐diamidino‐2‐phenylindole dihydrochloride (DAPI) for nuclear staining. Signals were captured using digital microscope (VH‐8000; Keyence, Osaka, Japan). Percentage of apoptotic cells were evaluated in five randomly selected fields (×100), and data are presented as mean value ± SD.