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. 2018 Oct 3;315(6):F1822–F1832. doi: 10.1152/ajprenal.00402.2018

Fig. 5.

Fig. 5.

Increased Smad2/3 expression in proximal tubule (PT)-Dicer knockout (KO) kidney may contribute to enhanced renal interstitial fibrosis in diabetic and unilateral ureteral obstruction (UUO) nephropathy. A: representative immunoblots of Smad2/3 in mouse kidney. C57BL/6 mice were subjected to either sham operation or UUO surgery. Mice were euthanized at 4 days (UUO4d), 1 wk (UUO1w), or 2 wk (UUO2w) after surgery, and left kidneys were collected for immunoblotting analyses of Smad2/3. Cyclophilin B was used as loading control. Representative immunoblots are presented. The results showed a time-dependent increase of Smad2/3 in kidney cortex following UUO. B: representative immunoblots and densitometry quantification of Smad2/3 levels. PT-Dicer wild-type (WT) and PT-Dicer KO mice were subjected to either sham operation or UUO surgery. Mice were euthanized at 1 wk after surgery, and left kidneys were collected for immunoblot analysis of Smad2/3 expression and densitometry quantification. The results showed the significantly increased Smad2/3 in kidney cortex in both PT-Dicer WT and PT-Dicer KO mice subjected to UUO compared with sham controls. The levels of Smad2/3 were significantly higher in PT-Dicer KO mice than in PT-Dicer WT mice after UUO. n ≥ 10. *P < 0.05, vs WT-sham; #P < 0.05, vs. PT-Dicer WT UUO. Cyclophilin B was used as loading control. C: representative immunoblots and densitometry quantification of Smad2/3 levels. Eight-week-old PT-Dicer WT and PT-Dicer KO mice were treated with streptozotocin (STZ) or without STZ (Control, CT) and examined after 16 wk. The results showed that Smad2/3 levels were significantly elevated in PT-Dicer KO mice compared with PT-Dicer WT mice in both control and diabetic mice. n ≥ 7. *P < 0.05, vs. WT-CT; #P < 0.05, vs. PT-Dicer WT diabetic mice. Cyclophilin B was used as loading control. D: representative immunoblot of Smad2/3. Left kidneys from 6-mo-old PT-Dicer WT and PT-Dicer KO mice were collected for immunoblot analyses. The results showed the remarkably increased Smad2/3 levels in PT-Dicer KO kidney cortex than in WT cortex. Cyclophilin B was used as loading control. E: Smad2 and Smad3 mRNA levels in the renal cortex in 6-mo-old PT-Dicer KO mice by real-time PCR; n = 5. No significant difference in Smad2/3 mRNA levels was found between WT and PT-Dicer KO mice under the control condition. F: representative images of Masson Trichrome staining of kidney samples from 7-mo-old PT-Dicer WT and PT-Dicer KO mice showing obvious deposition and accumulation of collagen in PT-Dicer KO mice compared with PT-Dicer WT mice; n = 6. Scale bar: 50 µm.