Fig. 6.

Targeting relationship between miR-665 and IL6 and their effects on ASCs. a qRT-PCR: expression of IL6 in normal/induced ASCs over time. b qRT-PCR: expression of IL6 in induced ASCs after agomiR-665, antagomiR-665, pcDNA3.1-IL6, or sh-IL6 transfection. c Predicted binding site between miR-556 and wt/mut IL6 3′UTR. d Dual-luciferase assay: luciferase activity in wt IL6 3′UTR + agomiR-665 group was significantly lower than that in wt IL6 3′UTR + agomiR-NC group. e qRT-PCR: expression of Runx2, Osterix, and Osteocalcin after transfection. f Western blot analysis of Runx2, Osterix, and Osteocalcin after transfection. g Alizarin red staining (21 days) and ALP staining (14 days): changes of calcium nodule formation and ALP activity after transfection. ^*P < 0.05, ^**P < 0.01, compared with NC group; ^#P < 0.05, ^##P < 0.01, compared with sh-NC group