Anti-Caspase 9/CASP9 Antibody Picoband®

Anti-Caspase-9 antibody, PA1595, Western blotting
All lanes: Anti Caspase-9 (PA1595) at 0.5ug/ml
Lane 1: SMMC Whole Cell Lysate at 40ug
Lane 2: MCF-7 Whole Cell Lysate at 40ug
Lane 3: CEM Whole Cell Lysate at 40ug
Lane 4: JURKAT Whole Cell Lysate at 40ug
Lane 5: RAJI Whole Cell Lysate at 40ug
Lane 6: HELA Whole Cell Lysate at 40ug
Predicted bind size: 35KD
Observed bind size: 35KD

Anti-Caspase-9 antibody, PA1595, IHC(P)
IHC(P): Human Tonsil Tissue

PPM-18 triggers apoptotic cell death in bladder cancer cells. (A) T24 and EJ cells were treated with the indicated concentration of PPM-18, and stained with annexin V-FITC and PI. The apoptotic effect of PPM-18 on T24 and EJ cells was measured by flow cytometry. (B) Western blotting showed the effect of PPM-18 on the expression of cleaved caspase-9, 3, and PARP in T24 and EJ cells. (C) T24 and EJ cells were exposed to 10 μM PPM-18 for 24 h, and stained with JC-1 fluorescent probe. The effect of PPM-18 on the alteration of mitochondria membrane potential was analyzed by flow cytometry. (D) Western blotting displayed the effect of PPM-18 on expression of BAX and BCL-2 in T24 and EJ cells. (E) and (F) The effect of Z-VAD-FMK on PPM-18 reduced the viability of T24 and EJ cells, or PPM-18 triggered apoptosis in T24 cells. Cells treated with 15 μM PPM-18 combined with or without 20 μM Z-VAD-FMK for 24 h, and cell viability and apoptosis were respectively measured by MTS assay and flow cytometry. Data are presented as the mean ± SD of at least three independent experiments. * p < 0.05, ** p < 0.01, and *** p < 0.001 vs. the control group, or vs. PPM-18+Z-VAD-FMK.
Index in PubMed under a CC BY license. PMID: 34305598

PPM-18 induces AMPK-dependent autophagy and apoptotic cell death in bladder cancer cells. (A) and (B) T24 and EJ cells were exposed to the indicated concentration of PPM-18 for 24 h, and the expression of phospho-AMPK, AMPK, phospho-mTORC1, phospho-P70S6K, phospho-PI3K, PI3K, phospho-AKT, and AKT were analyzed by western blot. (C) and (D) Western blot showed the effect of AMPK siRNA or 5 μM Compound C on the phosphorylation of AMPK, mTORC1, and P70S6K, and the expression of LC3B and cleaved caspase-9, 3 in PPM-18–treated T24 cells. (E) and (F) MTS assay and flow cytometry demonstrated that treatment with Compound C affected PPM-18–reduced viability of T24 cells and PPM-18–induced apoptosis in T24 cells. Data are presented as the mean ± SD of at least three independent experiments. *** p < 0.001 vs. the control group, or vs. PPM-18+Compund C.
Index in PubMed under a CC BY license. PMID: 34305598

PPM-18 stimulates autophagy that promotes apoptosis in bladder cancer cells. (A) T24 and EJ cells were treated with various concentrations of PPM-18 for 24 h. Then cells were harvested for western blotting to detect the expression of p62 and LC3B, two autophagy indicators. (B) The formation of autophagosomes in T24 and EJ cells treated with or without 10 μM PPM-18 for 18 h under the transmission electron microscope. Arrows indicated the autophagosomes containing intact and degraded cellular debris. Scale bar: left: 5.0 μm, middle: 2.0 μm, and right: 1.0 μm. (C) Western blot showed the effect of 5 mM 3-MA, a typical autophagy inhibitor, on the expression of p62, LC3B, and cleaved caspase-9, 3 in PPM-18–treated T24 and EJ cells. (D) and (E) T24 and EJ cells were treated with 15 μM PPM-18 combined with or without 5 mM 3-MA, and the effect of 3-MA on PPM-18–reduced viability or PPM-18–triggered apoptosis in T24 cells and EJ cells were measured by MTS assay and flow cytometry, respectively. (F) T24 and EJ cells were treated with 15 μM PPM-18 combined with or without 10 μM rapamycin, a typical autophagy agonist, and the effect of rapamycin on PPM-18–induced autophagy and apoptosis were evaluated by western blot. (G) The MTS assay demonstrated that rapamycin affected PPM-18–reduced viability in T24 and EJ cells. (H) Flow cytometry displayed the effect of rapamycin on PPM-18–triggered apoptosis in T24 and EJ cells. Data are presented as the mean ± SD of at least three independent experiments. ** p < 0.01 and *** p < 0.001 vs. the control group, or vs. PPM-18+3-MA. * p < 0.05 and *** p < 0.001 vs. PPM-18+Rapa.
Index in PubMed under a CC BY license. PMID: 34305598