Fig 2: Effect of salidroside (10 µM) on the levels of (a) p38, ERK, and JNK MAPKs and (b) Wnt/BMP signaling pathway. Analysis of protein levels was carried out by Western blotting using the osteo-induced hBM-MSC lysates at day 14 of differentiation with or without salidroside treatment. Western blot bands were densitometrically quantified and plotted as relative percentage of the untreated differentiated control group. Phosphorylation levels were measured via normalization of the phosphorylated (p-) protein against their total protein levels. ß-Actin was used as internal loading control. *p < 0.05 and **p < 0.01 vs. the untreated differentiated control group.

Fig 3: Effect of salidroside (10 µM) on the levels of (a) p38, ERK, and JNK MAPKs and (b) AMPK. Phosphorylated (p-) and total protein levels analyzed by Western blotting using adipo-induced hBM-MSC lysates at day 14 of differentiation. (c) Time-dependent p-AMPK levels were obtained at the designated times from adipo-induced compound C-treated hBM-MSCs lysates with or without salidroside (10 µM). Compound C was used as an inhibitor of AMPK phosphorylation. Western blot bands were densitometrically quantified and plotted as relative percentage of the untreated differentiated control group. Phosphorylation levels were measured via normalization of the phosphorylated (p-) protein against their total protein levels. ß-Actin was used as internal loading control. *p < 0.05 and **p < 0.01 vs. the untreated differentiated control group; #p < 0.05 and ##p < 0.01 vs. the differentiated control group without compound C and salidroside treatment.

Fig 4: miR-27a blocked JNK and Wnt/ß-catenin signaling pathways. A and C, Expressions of main proteins in JNK signaling, and B and D, expressions of main proteins in Wnt/ß-catenin signaling were measured after human mitral valve interstitial cells were transfected with miR-27a mimic/inhibitor and exposed to TNF-a. Data are reported as means±SD (n=3). ***P<0.001 vs the indicated group (ANOVA).

Fig 5: miR-27a blocked JNK and Wnt/ß-catenin signaling pathways in a NELL-1-dependent manner. A and C, Expressions of main proteins in JNK signaling, and (B and D) expressions of main proteins in Wnt/ß-catenin signaling were measured, after human mitral valve interstitial cells were co-transfected with miR-27a mimic and si-NELL-1, and were exposed to TNF-a. Data are reported as means±SD (n=3). ***P<0.001 vs the indicated group (ANOVA).