Fig 1: Changes in the transcript expression levels of the selected inflammation-associated genes (COX2 and NF?B), proto-oncogene (CTNNB1), autophagy-associated gene (CTSB), and apoptosis-associated genes (BCL2, CASP3, CASP7, CASP8, and CASP9). SKOV-3 cells were cultured with 0.1% (v/v) dimethyl sulfoxide (DMSO) alone (control) or DMSO containing a-mangostin (7.309 µM), apigenin (18.502 µM), or doxorubicin (0.431 µM) for 24 hr. Data are shown as the mean ± 1SD, derived from three independent repeats. * and ** represent a significant difference between the control and treated cells in each group at p < 0.05 or p<0.01, respectively.
Fig 2: Effects on ER stress-associated apoptosis in AGEs-treated SH-SY5Y cells. Cells were pretreated with diosmetin (DIOS; 10 µmol/L) or pioglitazone (PIO; 5 µmol/L) for 1 h, then exposed to 200 µg/mL AGEs for another 24 h. T0070907 (5 µmol/L) was added 1 h before diosmetin or pioglitazone were stimulated. (A) Representative photographs of Western blot analysis for Bax and Bcl-2. The ratio of the relative intensities of Bax to Bcl-2 (Bax/Bcl-2) was shown. (B) Analysis of caspase-12,-9-, and -3-like activity. The caspases-12-like activity in cell lysates was quantified by fluorescent detection of cleavage of substrate ATAD-AFC. Caspase-9 and caspase-3-like activities were measured against the colorimetric substrates Ac-LEHD-pNA and Ac-DEVD-pNA, respectively. (C) Apoptosis was determined by measuring cytoplasmic histone-associated DNA fragments using the cell death detection ELISAplus kit. The results are shown as the mean ± SD of five independent experiments (n = 5), each performed in triplicate. a p < 0.05 and b p < 0.01 compared to the data from untreated control group (control). c p < 0.05 and d p < 0.01 compared to the data from cells cultured under AGEs without any treatment.
Fig 3: Suppression of LBX2-AS1 inhibits proliferation and increases apoptosis of colorectal cancer cells. (A) LBX2-AS1 was silenced in HT29 and SW620 colorectal cancer cells following transfection with si-LBX2-AS1#1 or #2. (B) Cell Counting Kit-8 assay results showed that cell viability was inhibited in colorectal cancer cells transfected with si-LBX2-AS1. (C) The inhibitory effect of silencing LBX2-AS1 on colony formation was investigated in two colorectal cancer cells. (D) Flow cytometry assay results confirmed that the apoptosis of colorectal cancer cells was induced by si-LBX2-AS1. (E) Higher caspase-3 and caspase-9 expression levels were detected in colorectal cancer cells transfected with si-LBX2-AS1. **P<0.01 vs. si-NC. LBX2-AS1, LBX2 antisense RNA 1; si-, small interfering RNA; NC, negative control.
Fig 4: Analysis of cell viability and caspase-3 and caspase-9 activities after treatment of A549 and Hu02 cells with different concentrations of fraction 21 of P. persicus venom. (A) The dose-dependent effect of fraction 21 on the cell viability of A549 and Hu02 cell lines. (B) The caspase-3 and caspase-9 activities of the A549 and Hu02 cells after treatment with different concentrations of fraction 21. Relative caspase-3 and caspase-9 activities were estimated as a ratio of the cells treated with fraction 21 to untreated cell (control). (C) Effects of different concentrations of fraction 21 on the percentage of LDH released in A549 and Hu02 cells 24 hours after incubation. All data were presented as mean ± S.D (n = 3). Statistical significance versus control (*p < 0.05; **p < 0.01; ***p < 0.001).
Fig 5: Poricoic acid A (PAA) treatment induced apoptotic cell death. A, Flow cytometry analysis of Annexin V-positive cells after treatment with different concentrations of PAA for 24 h. B, Colorimetric assay of caspase-3, caspase-8, and caspase-9 activities using total cell lysates from cells treated with different concentrations of PAA for 6 h. C, Western blot analysis of caspase-3, cleaved caspase-3, Bax, Bcl2, and GAPDH protein levels in the absence or presence of 80 µg/mL PAA. Data are reported as means±SD of 3 independent experiments. *P<0.05, ***P<0.001 compared to Control (0 µg/mL) (two-tailed Student's t-test).
Supplier Page from Abcam for Caspase 9 Assay Kit (Colorimetric)