Bcl 2 sc 492

RPMI 1640 medium, gentamicin, trypsin, phosphate-buffered saline (PBS) and fetal bovine serum (FBS) were acquired from Gibco (Invitrogen, Carlsbad, CA, USA). Isolated compounds 7-hydroxy-3,4-dihydrocadalene (>98.0%), 7-(phenylcarbamate)-3,4-dihydrocadalene (77% yield) and 7-(phenylcarbamate)-cadalene (79% yield) were generously provided by Dr. Guillermo Delgado Lamas (Instituto de Química, UNAM). Their isolation or preparation and structural characterization are described by Egas et al. (2017) (link).
Dimethyl Sulphoxide (DMSO), H₂O₂, 2′,7′-Dichlorofluorescein Diacetate (DCFH-DA), 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), Bovine Serum Albumin (BSA), 2-thiobarbituric acid, antimycin A, oligomycin, rotenone, RIPA buffer, phosphatase inhibitor cocktails, bicinchoninic acid reagent, Tris Buffered Saline-Tween 20, GAPDH antibody (G-8795), caspase 3 (sc-7148) and Bcl-2 (sc-492) antibodies were purchased from Sigma-Aldrich (St. Louis, MO, USA). Protease inhibitor cocktail from Roche (Lewes, UK). Polyvinylidene difluoride (PVDF) membranes from GE Healthcare (Buckinghamshire, UK).

Based on Kim et al.’s paper [12 (link)], HCT116 and SW480 cells (1 × 10⁶ cells/mL) were exposed to Morusin (0, 2.5, or 5 μM) of for 24 h, The supernatants were collected and quantified for protein concentration by using RC DC protein assay kit (Bio-Rad, Hercules, CA, USA), The protein samples (30 μg of protein from cells) were separated on 4–12% NuPAGE Bis–Tris gels (Novex, Carlsbad, CA, USA) and ECL transfer membrane for detection with antibodies for PARP (#9542), pro-caspase-3 (#9662), cleaved caspase-3 (#9664), c-Myc (Y69) and ZNF746 (LS-B8045), Bcl-2 (SC-492), Bcl-xL (SC-8392), and β-actin (A2228) (Sigma, St. Louis, MO, USA). Proteins were detected using the ECL system (Amersham Pharmacia Biotech INC, District of Columbia, MO, USA). Densitometric analysis was performed using ImageJ software.