Ho 1 sc 10789

Pc was purchased from Sigma-Aldrich, Inc. (St. Louis, MO, USA). Go6976, was purchased from Calbiochem Company (San Diego, CA, USA). HO-1 and control siRNAs were obtained from Santa Cruz Biotechnology (Dallas, TX, USA) and antibodies to p-PKC α/β II (#9375), p-Akt (#2971), PARP-1 (#9542), p-p38 (#9212), cleaved caspase-3 (#9661), and β-actin (#4967) were obtained from Cell Signaling Technology (Beverly, MA, USA). p-Nrf-2 (ab76026) was purchased from Abcam (Cambridge, UK). Nrf-2 (sc-722), histone-H1 (sc-393358), procaspase-3 (H-277), and HO-1 (sc-10789) antibodies were purchased from Santa Cruz Biotechnology. All other reagents were purchased from Sigma-Aldrich unless otherwise indicated. Pc solutions was prepared in dimethyl sulfoxide (DMSO) and stored at −20 °C.

Immunohistochemical analysis was conducted as previously described by our research group (Rosa et al., 2017 (link)), with some modifications. In brief, paraffin-embedded tissues were deparaffinized, rehydrated in descending graded alcohols, incubated for 15 min in methanol containing 3% H₂O₂ to block endogenous peroxidase activity, and then subjected to microwave antigen retrieval for 30 min in sodium citrate buffer (10 mM tri-sodium citrate dihydrate, 0.05% Tween-20, pH 6.0). After preincubation in Super Block (ScyTek Laboratories, Logan, UT, United States) for 10 min, sections were incubated overnight with rabbit polyclonal anti-Egr-1 antibody (sc-110, Santa Cruz Biotechnology, Dallas, TX, United States, dilution 1:100) or rabbit polyclonal anti- HO-1 antibody (HO-1, sc-10789, Santa Cruz Biotechnology, dilution 1:100) at 4°C in humid chamber, washed with PBS, incubated for 10 min at room temperature with UltraTek Anti-Polyvalent (ScyTek Laboratories), washed with PBS and then incubated with the UltraTek HRP (ScyTek Laboratories) according to the manufacturer’s instructions. The sections were then stained with 3-3-diaminobenzidine (ScyTek Laboratories) as chromogen to visualize the reaction product, and were finally counterstained with hematoxylin. Images were acquired with Nikon Eclipse Ni motorized microscope system at 10× and 40× magnification.