Figure 3.

Lysicamine induces necroptosis in TC cells in a ROS-independent Manner. (a) Live cells (AV−/7-AAD−) were decreased in all TC cell lines by Lysicamine; (b) the same treatment resulted in increased necrotic cells (AV−/7-AAD+) in all TC cell lines; and (c) had no effect on Caspase 3/7 activation in all TC cell lines. (d) Lysicamine increased ROS production only in HTH83 (e) in a concentration-dependent manner. (f) In HTH83, ROS inhibition by NAC did not increase live cells (AV−/7-AAD−) or reduce necrotic cells (AV−/7-AAD+). (g) Necrosome inhibition via NEC1 treatment reduced necrotic cells (AV−/7-AAD+) triggered by lysicamine treatment. In all experiments, cells were treated with lysicamine IC₅₀ or DMSO (control) for 48 h, stained, and analyzed via flow cytometry. Results were expressed as percentages compared to untreated controls. Data were plotted as mean ± SEM of at least three biological replicates in triplicate; significance was determined by one-way or two-way ANOVA multiple comparisons followed by Bonferroni test (post-test); * p < 0.05, ** p < 0.001, and *** p < 0.0001.