Fig 2: Caspase-3, caspase-8 and caspase-9 relative activities in livers collected during and the post assisted-feeding period. (A) Caspase-3, caspase-8 and caspase-9 relative activities in livers at specific time points (n = 14) during the assisted-feeding period (days 84, 89, 96) and the post-overfeeding period (days 104, 117, 125). Values are means ± SEM. Bars with different letters (a-d) significantly differ (p < 0.05). Groups sharing the same letter are not significantly different. (B) Caspase-3, caspase-8 and caspase-9 relative activities in livers at specific time points (days 104, 117, 125) between previously assisted-fed ducks (n = 14) and ad libitum control ducks (n = 8). Significant differences are indicated by ****p < 0.000; NS: not significant.

Fig 3: C. iphionoides ethanolic extract induces mitochondrial dysfunction and activates intrinsic apoptotic signaling in Hep3B cells. Hep3B cells were treated with increasing concentrations of C. iphionoides ethanolic extract (10, 25, 50, and 100 µg/mL) for 24 h. (a) Mitochondrial membrane potential was assessed using the JC-1 assay and expressed as a percentage of JC-1 aggregate-positive cells, as determined by flow cytometry. (b) Activated Bax expression, (c) caspase-9 activity, (d) cleaved caspase-3 expression, (e) cleaved PARP-1 levels, and (f) BCL-2 expression were quantified following treatment. Activated Bax, cleaved caspase-3, cleaved PARP-1, and BCL-2 were analyzed by flow cytometry and expressed as a percentage of positive cells, whereas caspase-9 activity was measured using a fluorometric assay and expressed as relative fluorescence units (RFU). Data are presented as mean ± SD of five technical replicates. Statistical significance was determined using one-way ANOVA followed by Tukey's multiple comparisons test. Asterisks indicate comparisons with the vehicle-treated control group (0.1% DMSO): **p < 0.01, ****p < 0.0001.