fluorogenic substrate for caspase-3 ac-devd-afc  (Santa Cruz Biotechnology)

Journal: Cell Death Discovery

Article Title: Curcumin induced oxidative stress causes autophagy and apoptosis in bovine leucocytes transformed by Theileria annulata

doi: 10.1038/s41420-019-0180-8

Figure Lengend Snippet: a Western blot presenting the cleaved forms of caspase 3, caspase 8 and PARP upon curcumin treatment at various concentrations (5, 10, and 20 µM) for 24 h. Change in the expression level of the IKKb, Bcl2, and p-Bad upon curcumin treatment at various concentrations (5, 10, and 20 µM) for 24 h, b fold change in the expression of IKKb, Bcl2, and p-Bad upon curcumin treatment after normalization of western blot data, c After curcumin treatment, the activities of caspase 3 and caspase 8 were recorded. The activity of caspase 3 and caspase 8 were shown in the presence and absence of their inhibitors after 20 µM curcumin treatment. d Western blot showing the amount of cytochrome c present in cytoplasm after 20 µM curcumin treatment at different time points (6 h and 12 h). N = 3. Data are presented as mean ± SD. * represents p < 0.05, ** represents p < 0.01, and *** represents p < 0.001, compared with the untreated group

Article Snippet: Caspase 3 fluorogenic substrate Ac (N-acetyl)-DEVD-AFC, caspase 3 inhibitor (Ac-DEVD-CHO), caspase 8 fluorogenic substrate Ac-IETD-AFC, caspase 8 inhibitor (Z-IETD-FMK) and antibodies against caspase 8 and cytochrome c were purchased from BD Pharmingen.

Techniques: Western Blot, Expressing, Activity Assay

Journal: Oncotarget

Article Title: MicroRNA-497 impairs the growth of chemoresistant neuroblastoma cells by targeting cell cycle, survival and vascular permeability genes

doi: 10.18632/oncotarget.7005

Figure Lengend Snippet: ( A ) Proliferation time course comparing miR-497 versus miR-Control (25 nM) reverse transfected in CHLA-90 and SK-N-AS cells (both non-MYCN amplified) or SK-N-BE(2) and LA1-5s cells (both MYCN amplified). ( B ) Representative images of nuclear morphology assessment at 96 h post-transfection with Hoechst staining in miR-Control and miR-497 (25 nM) reverse transfected NB cell lines. Arrowheads point at condensed or fragmented nuclei. ( C ) Quantification of apoptosis was performed from 4 representative images of 3 replicates per condition. ( D ) Caspase-3/7 activity assays and ( E ) representative Western blot of PARP protein at 72 h post-transfection. SK-N-BE(2) and LA1-5s cells were reverse transfected with 25 nM of miR-Control or miR-497. Data represent mean ± SEM of three independent experiments *, ** or *** indicated significant differences comparing miR-497 versus miR-Control at p < 0.05, p < 0.01 or p < 0.001, respectively.

Article Snippet: Assays were performed in triplicate using 15–25 μg of protein in the same lysis buffer supplemented with 10 mM DTT and 25 μM of the fluorogenic caspase-3 substrate Ac-DEVD-AFC (Calbiochem).

Techniques: Transfection, Amplification, Staining, Activity Assay, Western Blot