Fig 2: Activation of ß-catenin by ß-catenin S33Y diminishes the protective roles of salidroside against ADR injury in podocytes. (A-B) The transfection of ß-catenin S33Y in cultured podocytes successfully activates ß-catenin activity. (C) Quantitative analysis of Axin2 and Cyclin D1 as shown in (B). (D) The repression of salidroside on ß-catenin activity was counteracted by ß-catenin S33Y in ADR-treated podocytes. (E) Quantitative analysis of Western blots as shown in (D). (F) The inhibition of ß-catenin downstream target gene expressions by salidroside was re-activated by ß-catenin S33Y in ADR-treated podocytes. (G) Quantitative analysis of Western blots as shown in (F). (H) Activation of ß-catenin by ß-catenin S33Y reverses the effects of salidroside on podocin and nephrin expressions in ADR-injured podocytes. (I) Quantitative analysis of Western blots as shown in (H). (J) ß-catenin S33Y has no effects on podocin and nephrin expressions. (K) Quantitative analysis of Western blots as shown in (J). (L) Effects of salidroside on Axin2 and Cyclin D1 expressions in podocytes transfected with the plasmid bearing ß-catenin S33Y. Lamin A/C and Actin were used as loading controls. The results are the means ± SEM of three independent experiments. EV, empty vector; CON, control; ADR, adriamycin; Sal, salidroside; MMP7, matrix metalloproteinase 7; AGT, angiotensinogen; PAI-1, plasminogen activator inhibitor-1. *P < 0.05; **P < 0.01 and ***P < 0.001 vs CON; # P < 0.05; ## P < 0.01 and ### P < 0.001 vs ADR; ^P < 0.05; ^^P < 0.01 and ^^^P < 0.001 vs ADR + Sal

Fig 3: Salidroside administration blocks renal ß-catenin activation induced by ADR. (A) Immunohistochemical analysis showing ß-catenin protein expression and localization in the kidney. Representative micrographs were shown. Scale bar = 20 µm. (B) Western blot analysis showing renal ß-catenin protein abundance in the nucleus and cytoplasm. Nucleus and cytoplasm lysates were distinguished using antibodies against Lamin A/C and Tubulin respectively (left panel). Lamin A/C and Actin were used as loading controls. (C) Quantitative analysis of ß-catenin protein levels as shown in (B). (D) Axin2 and Cyclin D1 expressions were examined. (E) Western blot analysis showing the effects of salidroside on ß-catenin downstream target gene expression. (F) Quantitative analysis of protein levels as shown in (E). Lamin A/C and Actin were used as loading controls. The results are the means ± SEM of three independent experiments. **P < 0.01; ***P < 0.001 vs CON; # P < 0.05; ## P < 0.01, ### P < 0.001 vs ADR alone. n = 5. ADR, adriamycin; AGT, angiotensinogen; CON, control; MMP7, matrix metalloproteinase 7; PAI-1, plasminogen activator inhibitor-1; Sal, salidroside