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. 2021 May 21;11:10676. doi: 10.1038/s41598-021-90115-8

Figure 3.

Figure 3

MSC mitochondrial transfer to iTregs occurs via TNT. (a) Mitochondrial quantity assay of iTregs was analyzed. At 3 weeks, media vs MSC co-culture iTreg cells were stained with NAO to quantify mt mass (n = 5). (b) Confocal imaging analysis for MitoTracker transfer. MSC were stained with CFSE (green) and MitoTracker Far Red (red), iTreg were stained with Hoechst (blue) and then co-cultured for 24–36 h (n = 5). (c) Mitochondria DNA copy number was assessed from IL-2/media or MSC co-culture expanded iTregs at day 21 (n = 6). (d) MSC mt transfer analysis. MSC were stained with CFSE (green) and MitoTracker Far Red (red), iTreg were stained Hoechst (blue) and cells were co-cultured for 24–36 h. Live cell images were collected by confocal microscopy. Data are representative of three independent experiments. (b,d) Analysis of recorded images was performed using Zen software Blue edition, (2011; version 2.0.14283.302). https://www.zeiss.com/microscopy/us/products/microscope-software/zen.html **p < 0.01, ***p < 0.001, ****p < 0.0001 paired t test. Scale bars: 10 μm. See also Supplementary Fig. S3.