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. 2019 Jan 18;27(3):571–583. doi: 10.1016/j.ymthe.2019.01.008

Figure 5.

Figure 5

Evidence of Exo/miR-29 in Kidney

Mice were injected with exosomes carrying either miR-control or miR-29. (A and B) Total RNA was isolated from (A) serum exosomes and (B) whole kidney of sham plus Exo/miR-ctrl (sham), sham plus Exo/miR29 (miR29), UUO plus Exo/miR-ctrl (UUO), and UUO plus Exo/miR-29 mice (UUO/29). The expressions of miR-29a-3p, miR-29b-3p, and miR-29c-3p were assayed by real-time qPCR. The bar graphs show miR expression of each group of mice compared with levels in control mice (represented by a line at 1-fold). (A) Bars represent mean ± SE; n = 6/group. *p < 0.05 versus sham; #p < 0.05 versus UUO. Results are normalized to miR103a. (B) Bars represent mean ± SE; n = 6/group. *p < 0.05 versus sham; #p < 0.05 versus UUO; &p < 0.05 versus miR29. Results are normalized to U6. (C) Mice were injected in the left TA with Exo/miR-29 labeled with 1 μmol/L fluorescent lipophilic tracer DiR at the same time as UUO surgery. The injection was repeated once per week. Shown are representative fluorescent kidney images at 14 days that were acquired using a Bruker Small Animal Optical Imaging System. Panels from left to right: sham operated with no exosome injection, UUO mouse with exosome injection, and sham-operated mouse with exosome injection. The bar graph shows fluorescence intensities of the kidneys. The original whole picture was shown in Figure S1 (bars: mean ± SE; n = 3/group; *p < 0.05, UUO versus sham mouse; #p < 0.05, UUO mouse obstructed kidney versus UUO mouse unobstructed kidney). (D) Circulation exosomes from different group mice were solubilized in Laemmli sample buffer, and fibrosis-related proteins TGF-β1, TGF-β3, and YY1 were analyzed by western blot. All blots were also probed for GAPDH, and all protein band densities have been normalized to their corresponding GAPDH loading control. The bar graph shows the fold change of each protein band compared with levels in control plus Exo/miR-ctrl (represented by a line at 1-fold) (bars: mean ± SE; n = 9/group; *p < 0.05 versus UUO; #p < 0.05 versus sham).