Extended Data Fig. 5: LC-EVs transport defective mitochondria to CD206hi tMacs.
From: An extracellular vesicle-mediated mitochondrial transfer network critical for testosterone synthesis
a, Schematic diagram depicts experimental design. Adult Cd11bDTR mice were weekly injected with vehicle (vehicle) or Diphtheria Toxin (DT) for a total of 3 consecutive injections, and testes were collected for analysis 7 days after the last injection. b, Representative confocal images of the testes. The testes sections were stained with IBA1 (n = 6 mice per group). Scale bars, 20 μm. c, Quantification of the number of tMacs between the 2 groups (n = 6 mice per group). d, Intratesticular testosterone levels of mice between the 2 groups (n = 6 mice per group). e, Schematic diagram depicts experimental design. Adult Cyp17a1Cre; R26tdTomato mice were unilaterally injected liposome-clodronate (CL2DMP) into the testis to deplete tMacs, while the contralateral testis was injected with control liposome (Con). The analyses were performed at indicated time points. f, Representative images of testis stained with IBA1 21 days after liposome injection (n = 5 mice per group). Scale bars, 50 μm. g, The number of tMacs at 21 days after liposome injection between the 2 groups (n = 5 mice per group). h, Intratesticular testosterone levels of mice at the indicated times after liposome injection between the 2 groups (n = 4 mice per group). i, The gating strategy for sorting CD206hi tMacs and MHCIIhi tMacs from Cyp17a1Cre; R26tdTomato; Cx3cr1GFP mice. j, Representative images of testis from Cyp17a1Cre; R26tdTomato mice stained with CD206 and MHCII (n = 5 mice). Scale bars, 5 μm. Inset shows boxed regions that are magnified. Scale bars, 2 μm. k, Percentage of CD206hi tMacs and MHCIIhi tMacs containing LC-EVs (n = 5 mice). l, Representative fluorescence micrographs of WT mice stained with CD206, MHCII, and CYP17A1 to detect the distance between LCs and tMacs subsets (n = 3 mice). Scale bars, 20 μm. m, Distance between LCs and CD206hi tMacs, MHCIIhi tMacs (n = 50 LCs from 3 mice). Data are represented by box plots (c,d,g) or mean ± SEM (k,m). Box plots: the box bounds depict the first quartile to the third quartile, with the middle line indicating the median and the whiskers extending to the minimum and maximum values. Significances were determined by Two-tailed Student’s t-test (c,d,g), Two-tailed paired t-test (h), Two-tailed Welch’s t-test (k) or Two-tailed Mann–Whitney U-test (m). Schematics in a,e created in BioRender. Xia, K. (2026) https://biorender.com/k1crpuu. Source numerical data are provided.
