Fig. 7: Post-NRC starvation induces Sit downregulation and moderate autophagy and thus precocious steroidogenesis.

A Heatmap represents expression profiles of the seven genes that were exclusively downregulated in post-NRC ST larvae. Numbers in the box represent log₂ expression values (zero-centered) that resulted from the integrated data analysis of the indicated genes. B, C qPCR analysis of sit was performed in RGs of the feeding (n = 3), pre-NRC ST (n = 3), post-NRC ST (n = 3) larvae or feeding larvae at different developmental points, including 96 h (n = 3), EW (n = 3), and LW (n = 3). Different lowercase letters represent statistical significance by one-way ANOVA. D, E The projected confocal z-stack showing that loss of sit in the PG at 96 h E, but not 84 h D, caused accumulation of lipid droplets. F The projected confocal z-stack showing that lipid droplets visualized by Nile Red staining (Red) in w¹¹¹⁸ PG cells at four developmental points. Magnified images of the indicated areas (yellow dotted box) are shown. n = 9 (96 h), 12 (108 h), 5 (EW), 5 (WPP). G Quantification of lipid droplets in the indicated time points shown in (F). Different lowercase letters represent statistical significance by one-way ANOVA. H The projected confocal z-stack showing lipid droplets visualized by Nile Red staining in w¹¹¹⁸ PG cells of larvae after 0 h, 12 h, and 24 h starvation from 84 h AEL or at WPP. Magnified images of the indicated areas (yellow dotted box) are shown. n = 9 (84 h), 8 (84 h ST + 12 h), 9 (84 h ST + 24 h), 7 (WPP). I Quantification of lipid droplets in the indicated time points shown in (H). Different lowercase letters represent statistical significance by one-way ANOVA. J Developmental timing and body size of Atg1 mutant (Atg1^−/−) larvae under feeding (n = 3) and starvation (n = 3) conditions. K, L Body size of Atg1 mutant (Atg1^−/−) under feeding and starvation conditions. Atg1^−/−: n = 47, Atg1^−/− ST: n = 9. Two-tailed paired t test for two-column comparison. ***p < 0.001. M Developmental timing of tub-Gal80^ts phm > w¹¹¹⁸ (n = 17), tub-Gal80^ts phm>sit (n = 13), tub-Gal80^ts phm>Atg1-RNAi (n = 7) and tub-Gal80^ts phm>sit+Atg1-RNAi (n = 16) under post-NRC ST conditions. All Data are presented as mean ± SD. The genotypes and statistical data are provided in Supplementary Data 1.