apoalert caspase 3 activity kit  (TaKaRa)

Journal: iScience

Article Title: Prenatal bisphenol A exposure reprograms SF1-lactylation pathways to promote endometriosis susceptibility

doi: 10.1016/j.isci.2026.115608

Figure Lengend Snippet: BPA exposure induces mitochondrial dysfunction, mitigated by NaOx or AC45594 HESC were treated with CTL or 50 nM BPA for 3 days, followed by 3-day treatment with vehicle, NaOx (20 mM), or AC45594 (5 μM). (A) Intracellular ATP. (B) Mitochondrial DNA copy number. (C) Mitochondrial membrane potential (ΔΨm). (D) Representative TMRE images. (E) Caspase-3 activity. (F) Apoptosis rate. n means number of independent repeats, n = 5. ∗, p < 0.05 vs. CTL(3d)+CTL(3d)/VEH; ¶, p < 0.05 vs. BPA(3d) + CTL(3d)/VEH. Data are presented as mean ± SD.

Article Snippet: ApoAlert Caspase-3 Activity Kit , Clontech , Cat# 630216.

Techniques: Membrane, Activity Assay

Journal: Cells

Article Title: Involvement of TRPA1 in Necrosis of Melanoma Cells via Phospholipase D1

doi: 10.3390/cells15090760

Figure Lengend Snippet: Transient receptor potential ankyrin 1 (TRPA1) activation induces necrosis in melanoma cells. ( a ) Effect of extracellular pH on melanoma cell viability. The cells were cultured at pH 5.4 (gray circles), 7.4 (open circles), and 8.1 (closed circles) for 3 days. Cell viability was assessed by MTT assay. ( b ) The cells were cultured with (closed circle) or without (open circle) 2 μM allyl isothiocyanate (AITC). Cell viability was assessed by MTT assay. ( c ) Dose-dependent effect of AITC on melanoma cell viability. The cells were treated with the indicated concentrations of AITC for 3 days, and cell viability was assessed by MTT assay. ( d , e ) Apoptosis (Annexin V) and necrosis (propidium iodide) in AITC-treated cells were detected using flow cytometry. Representative images of the flow cytometry analysis are shown ( d ), and the necrosis rate (positivity for propidium iodide) was calculated in the presence (closed circle) and absence (opened circle) of AITC. ( e ). Data are presented as means ± SEM of three independent experiments and compared using two-way ANOVA with Tukey’s honestly significant difference test for post hoc multiple comparison test ( a – c , e ). N.S., not significant, * p < 0.05, compared with pH 7.4 ( a ), untreated cells ( b , e ) or 0 μM ( c ).

Article Snippet: Annexin V and propidium iodide staining was performed with ApoAlert Annexin V-FITC Apoptosis Kit (TaKaRa Bio, Inc.).

Techniques: Activation Assay, Cell Culture, MTT Assay, Flow Cytometry, Comparison

Journal: Blood

Article Title: Withaferin A inhibits EBV-driven lymphomagenesis through multiple mechanisms, including EBNA1 degradation

doi: 10.1182/blood.2025029771

Figure Lengend Snippet: WA induces apoptosis in EBV + B-NHL cell lines. (A) Dose-response curves for EBV − (Akata 4E3) and EBV + (Akata BX1) cells (mean ± standard deviation of percentage of live cells relative to untreated). (B-D) Quantification of live cell numbers, dead cells (percentage of total), and caspase-3 activity shown as individual data points (n = 6). Statistical analysis for panels B-D: 2-way analysis of variance (ANOVA; factors: EBV status and dose) with Dunnett post hoc test comparing each dose to vehicle within EBV + and EBV − groups. ∗ P < .05; ∗∗ P < .01; ∗∗∗ P < .001; ∗∗∗∗ P < .0001. N/A, not available; ns, not significant; Veh, vehicle.

Article Snippet: Caspase-3 activity was determined using the ApoAlert Caspase-3 Fluorescence Assay kit (Takara Bio, 630215).

Techniques: Standard Deviation, Activity Assay

Journal: Blood

Article Title: Withaferin A inhibits EBV-driven lymphomagenesis through multiple mechanisms, including EBNA1 degradation

doi: 10.1182/blood.2025029771

Figure Lengend Snippet: WA increases survival of murine EBV + B-NHLs. (A) Spleen weights from mice treated with vehicle or WA (1, 5, or 10 mg/kg) on Mondays, Wednesdays, and Fridays for 30 days, beginning 5 days after EBV-infected cell inoculation (n = 16 mice per group; pooled from 4 independent experiments with unique cord blood donors). Statistical significance determined using 1-way ANOVA with the Dunnett post hoc test, comparing the vehicle, 5 mg/kg, and 10 mg/kg groups to the 1 mg/kg group. (B) EBV genome copy number from spleens in panel A, normalized to total DNA (n = 16 mice per group). Statistical significance determined using 1-way ANOVA with the Dunnett post hoc test, comparing each treatment group to vehicle. (C) Kaplan-Meier survival curves for vehicle- and WA-treated (5 mg/kg) cohorts (25 mice per group, pooled from 3 independent experiments with unique cord blood donors). Significance determined using the log-rank (Mantel-Cox) test. (D) Immunoblot analysis of spleen lysates (collected on the same dates for direct comparison) from the survival cohort, probing EBV proteins. (E) Tumor lysates from vehicle- and WA-treated mice (collected on the same dates for direct comparison) analyzed using SDS-PAGE and immunoblotting for viral proteins and apoptosis markers. (F) Quantification of Zta, phospho-p65, and cleaved caspase-3 protein levels from the immunoblots in panel E, normalized to actin, and expressed as fold change relative to vehicle tumors. Statistical significance determined using unpaired t tests. (G) Representative histopathology images of tumors from vehicle- and WA-treated cohorts. H&E staining (top) reveals extensive necrosis/apoptosis (≥50% of the tumor) in WA-treated tumors. Immunohistochemistry for CD20 (bottom) shows reduced and patchy CD20 staining in WA-treated tumors relative to vehicle controls. ∗ P < .05; ∗∗ P < .01; ∗∗∗ P < .001; ∗∗∗∗ P < .0001. Cl. Casp 3, caspase-3; H&E, hematoxylin and eosin; ns, not significant; Veh, vehicle.

Article Snippet: Caspase-3 activity was determined using the ApoAlert Caspase-3 Fluorescence Assay kit (Takara Bio, 630215).

Techniques: Infection, Western Blot, Comparison, SDS Page, Histopathology, Staining, Immunohistochemistry