Fig. 5
From: Thiostrepton induces apoptotic cell death at the level of BCL-2/CED-9 in C. elegans
Measuring the Generation of Reactive Oxygen Species Upon Thio Treatment. a Representative fluorescent image of wild-type worms stained with CellROX Green, comparing untreated (Blank), DMSO-treated, and 180 µM Thio-treated conditions. Thio-treated worms do not show significant changes in the GFP fluorescence intensity, indicating that Thio does not induce ROS formation. Scale bar = 20 μm. b Quantification of ROS levels based on GFP fluorescence intensity in wild-type worms following the treatments shown in a. Fluorescence was measured using ImageJ and statistically analyzed in GraphPad Prism. Bars represent mean ± SEM; ***p < 0.001 by one-way ANOVA. c Heatmap showing normalized GST-4::GFP fluorescence intensity in worms treated with Blank, DMSO, or Thio, measured every 2 h from 6 to 24 h post-treatment. Fluorescence intensity was quantified using ImageJ, normalized to background, and used to visualize dynamic GST-4 response across conditions. d Representative images of GST-4::GFP-expressing worms under each condition (Blank, DMSO, Thio) at each time point (6–24 h). These images illustrate the visual GST-4::GFP expression patterns corresponding to the quantified data shown in (c). e DIC images of adult germlines from daf-16(mu86) mutant worms treated with DMSO or Thio. Yellow arrowheads mark apoptotic cell corpses in the pachytene region. Scale bar = 20 μm. n > 50 germlines per condition. f Quantification of apoptotic germ cell corpses per gonad in wild-type and daf-16(mu86) mutant worms treated with Blank, DMSO, or Thio. Bars represent mean ± SEM. No statistically significant difference was detected between the wild type and daf-16(mu86) mutants. Statistical analysis was performed using one-way ANOVA; ns = not significant. n > 50 germlines per condition.
