Fig. 3: Fewer MERCs result in mitochondrial dysfunction and ER stress in aged SLCs. | Nature Communications

Fig. 3: Fewer MERCs result in mitochondrial dysfunction and ER stress in aged SLCs.

From: Nestin-dependent mitochondria-ER contacts define stem Leydig cell differentiation to attenuate male reproductive ageing

Fig. 3

a Flow cytometry of intracellular ROS level stained with DHE of Nestin-GFP + SLCs from different age groups in vivo. b Quantification of mean fluorescence intensity of DHE in (a). (n = 5 biological repeats for each group; One-way ANOVA). c qPCR analysis of relative mRNA expression of anti-oxidative genes of Nestin-GFP + SLCs from different age groups. (n = 5 biological repeats for each group; Multiple t tests). d, e Western Blot analysis and quantification of cellular anti-oxidative protein expression of Nestin-GFP + SLCs from different age groups. (n = 5 biological repeats for each group; Multiple t tests). f Representative immunostaining pictures of mitochondrial morphology of primary Nestin-GFP + SLCs from different age groups in vitro. Mitochondria are marked with Tom20. Scale bar, 20 μm. g A plot of Aspect Ratio (AR) against Form Factor (FF) showed that particles in primary Nestin-GFP + SLCs from different age groups in (f). Lower values of FF and AR, indicating small, circular mitochondria. h Representative TEM images of mitochondrial morphology of primary Nestin-GFP + SLCs from different age groups in vitro. Scale bar, 1 μm for original pictures and 500 nm for enlarged pictures. i Quantitative analysis of the average value of mitochondrial length (micrometers) in (h). (n = 5 biological repeats for each group; n = 126–184 mitochondria; One-way ANOVA). j qPCR analysis of relative mRNA expression of ER stress-related genes of primary Nestin-GFP + SLCs from different age groups in vitro. (n = 3 biological repeats for each group; Multiple t tests). k–o Western Blot analysis and quantification of ER stress-related proteins of primary Nestin-GFP + SLCs from different age groups in vitro. (n = 3 biological repeats for each group; Multiple t tests). Two-sided comparison; All data are mean ± SD; Error bars represent SDs. *p < 0.05, **p < 0.01, ***p < 0.001; Uncropped western blots and source data are provided as a Source Data file. See also Supplementary Figs. 3–5.

Back to article page