Rabbit polyclonal anti caspase 3

Mouse monoclonal anti-UHRF1 (1:2,000) antibody was engineered as previously described (1 (link)). Other antibodies used included rabbit polyclonal anti-HAUSP/USP7 (1:5,000; cat. no. ab4080, Abcam), mouse monoclonal anti-DNMT1 (1:5,000; cat. no. PTG-MAB0079, ProteoGenix), mouse monoclonal anti-ubiquitin (1:500; cat. no. 05-944, Sigma-Aldrich; Merck KGaA), mouse monoclonal eGFP (1:1,000; cat. no. 66,002-1-Ig, Proteintech Group, Inc.; and cat. no. A-11120, Thermo Fisher Scientific, Inc.), mouse monoclonal anti-GAPDH (1:5,000; cat. no. MAB374, Merck KGaA), mouse monoclonal anti-GFP (1:1,000; cat. no. 66002-1-Ig, Proteintech Group, Inc.), mouse monoclonal anti-p73 (1:500; cat. no. 558785, BD Biosciences), rabbit polyclonal anti-caspase-3 (1:1,000; cat. no. 9661, Cell Signaling Technology, Inc.), mouse monoclonal anti-BCL2 (1:1,000; cat. no. 05-826, Merck KGaA), mouse monoclonal anti-poly(ADP-ribose) polymerase (PARP; 1:1,000; cat. no. 51-6639GR, BD Biosciences) and rabbit polyclonal anti-BAX (1:1,000; cat. no. AB2930, Merck KGaA).

Immunohistochemical staining of activated caspase-3 and Bcl-2 was performed as previously described (Ibrahim et al., 2015 (link)). The heart sections were deparafinized, rehydrated and blocked in 3% hydrogen peroxide. Then, the sections were incubated with rabbit polyclonal anti-caspase-3 (1:1000 dilution; BD Biosciences, Le Pont-de-Claix, France) and mouse monoclonal anti-Bcl-2 (1:200 dilution, Santa Cruz Biotechnology) as primary antibodies. Diaminobenzidine (DAB) chromogen was added to visualize the immune reaction. Finally, the slides were counterstained in hematotoxylin and cover-slipped. Semi quantitative grading score for assessment of activated caspase-3 and Bcl-2 was carried out, in ten random high power fields, according to the percentage of immune stained cells in high power field (HPF) (40X). Score 0 = no staining; 1-positive staining in < 25% of cells/HPF; 2-positive staining in 25–50% of cells/HPF; 3-positive staining in >50% of cells/HPF.