MSM Enhances GH Signaling via the Jak2/STAT5b Pathway in Osteoblast-Like Cells and Osteoblast Differentiation through the Activation of STAT5b in MSCs
Figure 6
Methylsulfonylmethane (MSM)-enhanced GH signaling requires STAT5b activation in C3H10T1/2 cells.
(A) C3H10T1/2 cells were grown to 50% confluence and transfected with ON-TARGETplus SMARTpool siRNA targeting STAT5b or ON-TARGETplus Non-targeting siRNA using FuGene 6, according to the manufacturer’s instructions. 48 hours after transfection, cells were cultured with serum free osteogenic medium for 24 h and then cultured in osteogenic medium with 20 mM MSM for 24 h after the initiate osteoblast differentiation. Protein extracts (20 µg) were separated by 10% SDS-PAGE, and Western blots were performed. β-actin was used as a protein loading control. (B) The relative levels of protein were determined using densitometric analysis and normalized to the amount of β-actin. Data shown are representative of three independent experiments. Asterisks indicate a statistically significant increase by t-test (**p<0.01, ***p<0.001).