Fig 1: p53 mediates the regulation of lncCIRPIL on NMCMs injury induced by A/R.a Validation of p53 overexpression plasmids in NMCMs. n = 3. *P < 0.05 vs NC (negative control) by two-tailed Student’s t test. b Effect of p53 co-transfection on LDH release of lncCIRPIL overexpressing NMCMs subjected to A/R insult. n = 3. *P < 0.05 by one-way ANOVA followed by Tukey post hoc analysis. c Co-transfection of LncCIRPIL and p53 plasmids on caspase-3 activity in NMCMs subjected to A/R insult. n = 3. *P < 0.05 by one-way ANOVA followed by Tukey post hoc analysis. d. Co-transfection of LncCIRPIL and p53 plasmids on Bcl2/Bax ratio in NMCMs subjected to A/R insult. n = 3. *P < 0.05 by one-way ANOVA followed by Tukey post hoc analysis. e LDH release of NMCMs co-transfected with lncCIRPIL siRNA and p53 siRNA. n = 3. *P < 0.05 by one-way ANOVA followed by Tukey post hoc analysis. f Caspase-3 activity in NMCMs co-transfected with lncCIRPIL siRNA and p53 siRNA. n = 3. *P < 0.05 by one-way ANOVA followed by Tukey post hoc analysis. g Bcl2/Bax ratio in NMCMs subjected to A/R insult co-transfected with lncCIRPIL siRNA and p53 siRNA. n = 3. *P < 0.05 by one-way ANOVA followed by Tukey post hoc analysis.
Fig 2: Leptin rescues gallbladder cancer cells from gemcitabine-induced cell apoptosis.(A) The viability of leptin–treated GBC cells in response to gemcitabine (GEM). SNU-308 and RCB-1130 cells were treated with GEM alone or in combination treatment with leptin. Cell viability was assessed by the MTT assay (n = 3; ***P < 0.001, **P < 0.01, 2-tailed Student’s t test). (B) RCB-1130 cells were starved for 24 hours and then treated with 100 nM gemcitabine or a combination of gemcitabine and leptin (100 ng/mL) for 24 hours. Cell toxicity was measured by the LDH assay (n = 3; **P < 0.01, ***P < 0.001), 1-way ANOVA followed by Tukey’s multiple comparisons). (C) Leptin attenuates GEM-induced caspase-3 activation. Caspase-3 activity was detected in SNU-308 and RCB-1130 cells (n = 3; ***P < 0.001, 2-tailed Student’s t test). (D) SNU-308 and RCB-1130 cells were plated in 100 nM GEM or 100 nM GEM in combination with leptin and treated for 24 hours in microtiter plates. Cell apoptosis was analyzed by the TUNEL assay. Blue spots represent cell nuclei by DAPI staining, and green spots represent apoptotic bodies by TUNEL staining (n = 3, 1-way ANOVA followed by Tukey’s multiple comparisons). Scale bar: 100 μm.
Fig 3: Reduction in caspase-3 activity by compound 18 in SH-SY5Y cells. Cells were treated with 18 at 0, 20, and 40 µM for 24 h at 37° and then with H2O2 500 µM for 4 h. Data are presented as the Mean ± SEM of three different experiments carried out in quadruplicate. # p <0.001 vs. vehicle-treated control. ** p < 0.01 vs. only with H2O2-treated cells.
Fig 4: LINC00319 participates in OGD-induced cerebral ischemic injury. (A) Transfection efficacy of pcDNA3.1 (OE-LINC00319) or siRNA (si-LINC00319) in OGD-induced primary neurons as assessed using a reverse transcription-quantitative PCR assay. (B) Cell viability was detected in OGD-induced primary neurons transfected with OE or si-LINC00319 with a Cell Counting Kit-8 assay. (C) Caspase-3 activity was detected in OGD-induced primary neurons transfected with OE or si-LINC00319 with a Caspase-3 Fluorometric Assay. (D) Glucose uptake was measured in OGD-induced primary neurons transfected with OE or si-LINC00319 via the glucose oxidase-peroxidase method. (E) The expression of Caspase-3 was detected in OGD induced primary neurons transfected with OE or si-LINC00319 by western blotting assay (F). Apoptotic rate was measured in OGD-induced primary neurons transfected with OE or si-LINC00319 via Hoechst-33258 assay. *P<0.05, **P<0.01, ***P<0.001. Data are presented as the mean ± SD of three independent experimentsOGD, oxygen-glucose deprivation; siRNA, small interfering RNA; OE, overexpression.
Fig 5: Effects of lncCIRPIL on anoxia/reoxygenation(A/R) induced injury of NMCMs.a Validation of lncCIRPIL overexpression plasmid in NMCMs. n = 3. *P < 0.05 vs NC (negative control) by two-tailed Student’s t test. b Knockdown efficiency of shRNAs for lncCIRPIL in NMCMs. n = 3. *P < 0.05 vs NC by two-tailed Student’s t test. c Effects of lncCIRPIL on LDH release of NMCMs subjected to A/R insult. n = 4. *P < 0.05 by one-way ANOVA followed by Tukey post hoc analysis. d Effects of lncCIRPIL on cell apoptosis of NMCMs subjected to A/R insult by TUNEL staining. n = 18 slices from 3 independent experiments for each group. *P < 0.05 by one-way ANOVA followed by Tukey post hoc analysis; Scale bar = 50 μm. e Effects of lncCIRPIL on Bcl2/Bax ratio of NMCMs subjected to A/R insult. n = 4. *P < 0.05 by one-way ANOVA followed by Tukey post hoc analysis. f Effects of lncCIRPIL on caspase-3, caspase-8, caspase-9 activities of NMCMs subjected to A/R insult. n = 3. *P < 0.05 by one-way ANOVA followed by Tukey post hoc analysis.
Supplier Page from Abcam for Caspase-3 Assay Kit (Fluorometric)