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Fig. 4 | Arthritis Research & Therapy

Fig. 4

From: MicroRNA-15b shuttled by bone marrow mesenchymal stem cell-derived extracellular vesicles binds to WWP1 and promotes osteogenic differentiation

Fig. 4

KLF2 overexpression promotes osteogenic differentiation in BMSCs by suppressing the transcription activity of NF-κB. a The protein expression of KLF2 during osteoblasts differentiation assessed by Western blot analysis normalized to GAPDH. *p < 0.05 vs. PM. b KLF2 expression in BMSCs after transduction with oe-KLF2, sh-KLF2-1, or sh-KLF2-2. c, d ALP activity in BMSCs after the alteration of KLF2 (× 200). e The expression of Runx2, OPN, and OCN in BMSCs with osteogenic differentiation for 14 days after the alteration of KLF2 assessed by Western blot analysis normalized to GAPDH. *p < 0.05 vs. BMSCs transfected with sh-NC. #p < 0.05 vs. BMSCs transfected with oe-NC. f The mineralization of BMSCs determined using ARS staining after treatment with sh-NC, sh-KLF2, oe-NC, or oe-KLF2. *p < 0.05 vs. BMSCs treated with sh-NC. #p < 0.05 BMSCs treated with oe-NC (× 200). g The interaction of KLF2 with NF-κB detected by dual-luciferase reporter assay. *p < 0.05 vs. oe-NC. #p < 0.05 vs. p65 + oe-NC. h, i ALP activity in BMSCs after the alteration of KLF2 and NF-Κb (× 200). j The expression of Runx2, OPN, and OCN in BMSCs with osteogenic differentiation for 14 days after the alteration of KLF2 and NF-κB measured by Western blot analysis normalized to GAPDH. *p < 0.05 vs. BMSCs transduced with sh-KLF2. #p < 0.05 vs. BMSCs transduced with oe-KLF2. k The mineralization of BMSCs determined using ARS staining after treatment with sh-KLF2 + Tanshinone-I, sh-KLF2, oe-KLF2 + TNF-α, or oe-KLF2 (× 200). *p < 0.05 vs. BMSCs transduced with sh-KLF2. #p < 0.05 BMSCs transduced with oe-KLF2. The measurement data were shown as mean ± standard deviation. Data between the two groups were compared by independent sample t test. Comparisons among multiple groups were performed using one-way ANOVA, followed by Tukey’s post hoc test. The cell experiment was repeated three times

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