Fig. 3: A Nacα allele reduces its expression for cell death induction.

a, b Relative gene expression levels of Nacα/β and sug as determined by qRT-PCR. Second or early third instar larvae of the indicated genotypes were used. c–e Caspase-positive fat body cells. Fat bodies were dissected from the mid-third instar and stained for cDcp1, F-actin, and DNA (c). Arrows indicate cDcp1-positive cells. Quantification of nuclear diameter (d) and cell area (e) in cDcp1-negative (–) or -positive (+) fat body cells. Each plot against the cDcp1 mean fluorescence intensity per cell is shown on the right (d, e). f Formation of apoptotic bodies in Nacα mutant fat body. Arrows indicate cDcp1-positive dispersed puncta along the cell-cell boundaries. g Secondary necrosis in fat body cells. Fat bodies were dissected from the mid-third instar and stained for Annexin V-FITC, PI, and Hoechst. Cyan and yellow arrows indicate Annexin V-positive/PI-negative and Annexin V/PI double-positive cells, respectively. h, i Characterization of caspase-positive fat body cells. cDcp1-positive Nacα mutant fat body cells are positive for TUNEL staining (h) and GC3Ai (i), as indicated by arrows. High-magnification images of dashed areas are shown (g, h). Horizontal lines indicate the means of individual groups (d, e). Values of n indicate the number of nuclei or cells (d, e) from multiple animals. For appropriate panels, results are presented as the mean ± SD (a, b); n = 4 batches (b); one-way ANOVA with Tukey’s post hoc test (a), one-way ANOVA with Dunnett’s post hoc test (b), unpaired two-sided Mann–Whitney U-test (d, e). Scale bars: 50 μm (c, f–i).