Fig. 3: Autophagy is activated by MitoCa2+ to delay ISC aging.
a–i Quantification of the number of mCherryATG8 (a, g), lysotracker (b, h), Magic Red (c), DQ-BSA (d), autophagosomes (f), autolysosomes (e, f) and Ref(2)P-GFP (i) positive puncta in young ISCs (3 d) with indicated genotypes and conditions, respectively. Data show mean ± S.E.M (n = 102, 111, 91 cells in a, n = 71, 66, 63 cells in b, n = 130, 96 cells in c, n = 76, 68 cells in d, n = 100, 100, 102 cells in e, n = 100, 101, 100, 94 cells in f, n = 112, 123, 147, 163, 140, 124 cells in g, n = 101, 122, 120, 105, 122, 106 cells in h and n = 75, 70, 54 cells in i, respectively, from left to right) based on one-way ANOVA with Tukey’s multiple comparison test (a, b, e–i) and two-tailed unpaired t-test (c, d). ****P  <  0.0001. j Representative images and quantification of the number of autophagosomes and/or autolysosomes in young ISCs (3 d) with indicated genotypes by TEM. VM, visceral muscles; M, mitochondria; N, nucleus. ISCs and their nucleus were circled in light yellow and typical autophagosomes or autolysosomes were highlighted by arrowheads. Data show mean ± S.E.M (n = 20, 21 and 20 cells, from left to right) based on one-way ANOVA with Tukey’s multiple comparison test. *P  <  0.05, **P  <  0.01 and ****P  <  0.0001. Scale bar, 1 µm. k Quantification of clone numbers in the posterior midgut with indicated genotypes at different ages after treated or not with CQ for indicated time. Data show mean ± S.E.M (n = 13, 11, 10, 12, 13, 11, 11 and 14 guts, from left to right) based on one-way ANOVA with Tukey’s multiple comparison test. ****P  <  0.0001. l, m Quantification of the intensity of anti-γH2AvD signals in ISCs of aged flies (30 d) with indicated genotypes, respectively. Data show mean ± S.E.M (n = 8, 8, 9, 9 guts in l and n = 13, 9 guts in (m) from left to right) based on one-way ANOVA with Tukey’s multiple comparison test (l) and two-tailed unpaired t-test (m). ****P  <  0.0001. Each experiment was repeated for 3 times (a–m). See Supplementary Table 1 for genotypes. Source data are provided as a Source Data file.
