Fig. 5: MERCs-mediated calcium crosstalk between the ER and mitochondria involved in MitoCa2+-mediated autophagy in ISCs. | Nature Communications

Fig. 5: MERCs-mediated calcium crosstalk between the ER and mitochondria involved in MitoCa2+-mediated autophagy in ISCs.

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

Fig. 5

a Representative images and quantifications of the number and intensity of MAM-GFP positive vesicles (green) in young ISCs (3 d) with indicated genotypes. Typical ISCs (red) were circled in dashed lines. Data show mean ± S.E.M (n = 98, 107, 104, 121 and 107 cells, from left to right) based on one-way ANOVA with Tukey’s multiple comparison test. ****P  <  0.0001. Scale bars,5 μm. b Quantification of relative MitoCa2+ levels in young ISCs (3 d) with indicated genotypes. Data show mean ± S.E.M (n = 123, 136, 104, 108 and 111 cells, from left to right) based on one-way ANOVA with Tukey’s multiple comparison test. ****P  <  0.0001. c Quantification of the number of MAM-GFP positive vesicles in young ISCs (3 d) with indicated genotypes. Data show mean ± S.E.M (n = 129 and 127 cells, from left to right) based on two-tailed unpaired t-test. ****P  <  0.0001. d, e Representative TEM images (d) and quantifications (e) of MERCs in young ISCs (3 d) with indicated genotypes. Typical ISCs nucleus (yellow). Data show mean ± S.E.M (n = 8 and 8 sections, from left to right) based on two-tailed unpaired t-test (e). **P  <  0.01. Scale bars,2 μm (d). f Quantification of the number of MAM-GFP positive vesicles in young ISCs (3 d) with indicated genotypes. Data show mean ± S.E.M (n = 124, 114, 97, 100 and 95 cells, from left to right) based on one-way ANOVA with Tukey’s multiple comparison test. ****P  <  0.0001. g–j Quantification of the number of mCherryATG8a (g, i) and lysotracker (h, j) positive puncta in young ISCs (3 d) with indicated genotypes. Data show mean ± S.E.M (n = 69, 75, 85, 97 cells in g, n = 93, 81, 88, 81 cells in h, n = 131, 115, 102 cells in (i) and n = 90, 79, 75 cells in (j), from left to right) based on one-way ANOVA with Tukey’s multiple comparison test. ****P  <  0.0001. k Representative images and quantification of the number of autophagosomes in young ISCs (3 d) with indicated genotypes by anti-ATG8a staining (magenta). ISCs (green). Data show mean ± S.E.M (n = 76 and 66 cells, from left to right) based on two-tailed unpaired t-test. ****P  <  0.0001. Scale bars, 5 μm. Each experiment was repeated for 3 times (a–k). See Supplementary Table 1 for genotypes. Source data are provided as a Source Data file.

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