Fig. 4

MiR-617 inhibits DPSCs osteogenesis. a Western blot assay showed higher protein levels of ALP, RUNX2, and OSX in the NC group and miR-617 inhibitor group than mimics group and miR-617 inhibitor NC group respectively. GAPDH was the internal control. b Grayscale analyses. *P < 0.05 or **P < 0.01. c RT-PCR showed higher levels of ALP, RUNX2, and OSX in the NC group and inhibitor than mimics group and iNC group, respectively. d ALP staining in DPSCs following a 7-day osteogenesis with overexpression or knockdown of miR-617. e Histograms showed quantification of Alizarin red staining by spectrophotometry. f After 14 days of co-culture, upper: alizarin red staining showed that miR-617 mimics group generated more calcified nodules than the control group. MiR-617 inhibitor group generated more calcified nodules than the iNC group. Lower: mineralized nodules in different groups under the inverted microscope (OM: osteogenic medium). g Immunofluorescence showed positive expressions of ALP and OSX in DPSCs with overexpression or knockdown of miR-617. *P < 0.05, **P < 0.01