Fig. 1: Deficient MitoCa2+ uptake contributes to ISC dysfunction in aged Drosophila. | Nature Communications

Fig. 1: Deficient MitoCa2+ uptake contributes to ISC dysfunction in aged Drosophila.

From: Restoring calcium crosstalk between ER and mitochondria promotes intestinal stem cell rejuvenation through autophagy in aged Drosophila

Fig. 1

a Plot summary of calcium level in cytosol and mitochondria at different time points. A, amplitude. b Representative images and quantification of MitoCa2+ (green) and CytoCa2+ (red) in ISCs of young (3 d) and old animals (30 d) with indicated genotypes exposed or not to Ecc15 infection. Data show mean ± S.E.M (n = 95, 122, 99 and 173 cells, from left to right) based on two-way ANOVA with Tukey’s multiple comparison test. ****P  <  0.0001. Scale bar, 5 µm. c Representative images and quantification of relative MitoCa2+ levels in ISCs of young (3 d) or old animals (30 d) with indicated genotypes. MitoGCaMP3 (green), MitoDsRed (red). Data show mean ± S.E.M (n = 109, 110, 131, 104, 93 and 104 cells, from left to right) based on two-way ANOVA with Tukey’s multiple comparison test. ****P  <  0.0001. Scale bar, 5 µm. d Quantification of relative MitoCa2+ levels in aged ISCs (30 d) with indicated genotypes exposed or not to Ecc15 infection. Data show mean ± S.E.M (n = 133, 129, 108 and 126 cells, from left to right) based on two-way ANOVA with Tukey’s multiple comparison test. ****P  <  0.0001. e Quantification of the number of mis-differentiation ISCs with indicated genotypes at different ages. Data show mean ± S.E.M (n = 10, 8, 8 and 6 guts, from left to right) based on two-way ANOVA with Tukey’s multiple comparison test. ****P  <  0.0001. f Schematics for MARCM clone analysis. Main cell types (upper left), clonal formation (clone of GFP positive cells) and clonal maintenance (shedding GFP positive cells in the clone) (bottom) were shown as schematics. g Representative images and quantification of clone numbers in the posterior midgut with indicated genotypes at different ages. ISC clones (green) and nuclei (blue). Data show mean ± S.E.M (n = 7, 9, 10, 8, 7 and 7 guts, from left to right) based on two-way ANOVA with Tukey’s multiple comparison test. *P  <  0.05 and ****P  <  0.0001. Scale bar: 50 µm. Each experiment was repeated for 3 times (b–e, g). See Supplementary Table 1 for genotypes. Source data are provided as a Source Data file.

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