After a blocking incubation with 5% milk-TBST, the membranes were incubated with primary antibodies at a dilution of 1:1000 for the specific detection of caspase-3 (ab4051), cleaved caspase-3 (ab32042), caspase-9
(ab32539), cleaved caspase-9
(ab32539), PARP (ab32138), cleaved PARP (ab32138), p53 (ab131442), mdm2 (ab38618), p-p53(ab1431), p21 (ab109520), Bcl-2 (ab32124), Bax (ab32503), [gamma]-H2AX (ab11174), and [beta]-actin (ab8227, Abcam, USA) at 4[degrees]C overnight.
It involves the activation of caspase-9
which then triggers a cascade of events involving caspase-3 and caspase-7 activation8.
Membranes were blocked with 5% blotting-grade milk and incubated with primary antibodies (rabbit anti-cleaved caspase-9
or anti-cleaved caspase-3) at 4[degrees]C overnight.
The intrinsic pathway is begun by changes in mitochondrial permeability and the activation of caspase-9
The perfluorooctane sulfonate triggered the transactivation of Bax and activate caspase-3 and caspase-9
in zebrafish embryos .
Sui et al., "Acidic pH inhibits ATP depletion-induced tubular cell apoptosis by blocking caspase-9
activation in apoptosome," American Journal of Physiology Renal Physiology, vol.
Cell counting kit 8 (CCK-8) and caspase-3, caspase-8, and caspase-9
fluorescence metric assay kits were purchased from KeyGen BioTECH (Jiangsu, China).
The level of caspase-9
was evaluated significantly at I[C.sub.75] of FAA in MCF-7.
We found that FK866 and CHS828, two distinct NAMPT inhibitors, increased the TMZ-induced apoptosis and necrosis, enhanced the TMZ-induced caspase 1, caspase-3, and caspase-9
activities, and augmented the TMZ-induced oxidative stress in glioblastoma cells.
In addition, trifolin increased the levels of the death receptor involving the Fas/Fas ligand (FasL) and Fas-associated protein with the death domain (FADD), which consequently activated caspase-8, caspase-9
, caspase-3, and the proteolytic cleavage of poly (ADP-ribose) polymerase (PARP).
Caspase-3 is activated by the upstream caspase-8 and caspase-9
, and accordingly it serves as a convergence point for different signaling pathways .
In Western blot tests, after electrophoresis, the transferred PVDF membranes were incubated overnight with antibodies against Akt, phosphoAkt (Ser473), GSK-3[beta], and phospho-GSK-3[beta] (Ser9) (Cell Signaling Technology, MA, USA), B-cell lymphoma-2 (Bcl-2), Bax, or cleaved cysteine-dependent aspartate-directed proteases (caspase-3, caspase-6, or caspase-9
) (Abcam, CA, USA).