Figure 3.
Silent mating‐type information regulation 2 homolog 1 (SIRT1) inhibition induces intrinsic apoptotic signaling in adapted suspension cells (ASC). A, Adherent cells (AD) and ASC were plated at a density of 5 × 105 per well and cultured for 72 h with the indicated concentration of nicotinamide (NAM). Level of cleaved poly ADP ribose polymerase (PARP) as a marker of apoptosis was determined by immunoblot assay. B, Levels of cleaved PARP, caspase‐3, ‐8, and ‐9 were examined in SIRT1‐depleted cells using immunoblot assay. C, Level of death receptor signaling associated protein in SIRT1‐depleted cells was determined by immunoblot assays. D, Level of BCL‐2 family protein in SIRT1‐depleted cells was determined by immunoblot assays. E, Cells transfected with siControl and siSIRT1 were stained with JC‐1 dye and analyzed by flow cytometry. Bar diagram shows percentage of cells showing mitochondrial depolarization. *P < .05