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. 2022 Oct 9;12(16):7158–7179. doi: 10.7150/thno.78376

Figure 11.

Figure 11

PA-S14 attenuates renal fibrosis and tubular cell senescence in UUO mice. (A) Diagram shows the experimental design. Red arrows indicate the time points undergoing UUO and sacrifice, respectively. Green arrows indicate PA-S14 treatment (1.0 mg/kg body weight). (B) Quantitative analysis of tubular injury in different groups as indicated. Kidney sections were subjected to PAS staining. At least 10 randomly selected fields were evaluated under 400× magnification and results were averaged for each animal. ***P < 0.001 versus sham controls; ##P < 0.01versus UUO (n = 5). (C) Representative staining micrographs show Periodic acid-Schiff (PAS) and Sirius red staining. Paraffin sections were performed to PAS and Sirius red staining. Arrows indicate positive staining. Scale bar, 50 µm. (D) Graphical representations of the degree of kidney fibrotic lesions in different groups after quantitative determination of Sirius red staining intensity. ***P < 0.001 versus sham controls; ###P < 0.001 versus UUO (n = 5). (E-H) Representative western blot and quantitative data show the reduction of renal Fibronectin, Collagen I and α‐SMA expression after PA-S14 treatment. Numbers (1-3) indicate each individual animal in given group. **P < 0.01 versus sham controls; ##P < 0.01 versus UUO (n = 5). (I) Representative micrographs show the expression of active β-catenin, Fibronectin, α‐SMA, SA‐β‐gal activity, Klotho and KIM-1 in different groups as indicated. Frozen kidney sections were stained for active β-catenin and SA‐β‐gal activity. Paraffin sections were stained with antibodies against Fibronectin, α‐SMA, Klotho, and KIM-1, respectively. Arrows indicate positive staining. Scale bar, 50 µm. (J-N) Representative western blot and quantitative data showing renal expression of active β-catenin, p16INK4A, Klotho and KIM-1. Numbers (1-3) indicate each individual animal in a given group. **P < 0.01, ***P < 0.001 versus sham controls; ##P < 0.01, ###P < 0.001versus UUO (n = 5). (O) Quantitative determination of SA‐β‐gal activity staining in UUO kidney. ***P < 0.001 versus sham controls; ###P < 0.001 versus UUO (n = 5).