Figure 6

Immunofluorescent stains for presence of CD31⁺and vWF⁺cells in renal parenchyma after ischemia–reperfusion. Immunofluorescent (IF) stains for presence of CD31⁺ and von Willebrand Factor (vWF)⁺ cells in renal parenchyma at 72 hours after ischemia–reperfusion (IR) (n = 8). (A) to (E) IF staining (200×) of CD31⁺ cells (white) in renal parenchyma. Notably fewer CD31⁺ cells in the IR group (B) than in other groups (A, C, D, E). (D), (E) Merged image from double-staining (DiI + CD31) showing cellular elements of mixed red and yellow (red arrows) under higher magnifications ((B) and (D) are magnified images of (A) and (C), respectively), indicating implanted numerous doubly-stained cells (CD31⁺) in peri-tubular and interstitial areas. DAPI counter-staining for nuclei (blue). Scale bars at right lower corner = 50 μm. F) *P <0.0001 vs. other groups with different symbols; † vs. ‡ vs. ¶ vs. §, all P <0.05 for inter-group comparisons. (G) to (K) IF staining (200×) of vWF⁺ cells (white) in renal parenchyma. Notably fewer vWF⁺ cells in the IR group (H) than in other groups (G, I, J, K). (K) Merged image from double-staining (DiI + vWF) showing cellular elements of mixed red and yellow (red arrows) under higher magnifications ((F) and (H) are magnified images of (E) and (G), respectively), indicating implanted doubly-stained cells (vWF⁺) in peri-tubular and interstitial areas. DAPI counter-staining for nuclei (blue). Scale bars at right lower corner = 50 μm. (L) *P <0.0001 vs. other groups with different symbols; † vs. ‡ vs. ¶ vs. §, all P <0.05 for inter-group comparisons. All statistical analyses performed using one-way analysis of variance followed by Bonferroni multiple comparison post-hoc test. Different symbols (*, †, ‡, ¶, §) among different groups indicate significance at the 0.05 level. ADMSC, adipose-derived mesenchymal stem cells; CsA, cyclosporine A.