Further, the western blot assay revealed that the protein levels of ALP, Runx2
, and OPN were notably up-regulated in a time-dependent manner (P< 0.001, Figure 1D and E).
regulates the expression of bone-related proteins such as osteocalcin, osteopontin, as well as receptor activator of nuclear factor kappa-B ligand (RANKL), and alkaline phosphatase (ALP). Osterix is located downstream of Runx2
, and its main role is to increase the expression of osteopontin and ALP. Thus, VSMCs undergo phenotypic transformation and downregulation of expression of the contractile proteins smooth muscle (SM) 22 and alpha-smooth muscle actin (a-SMA).
No statistically significant differences were observed for the expression of the proinflamatory cytokines IL1A, IL6, IL8 and TNF and bone formation genes ALP, COL1 and RUNX2
in HPDL cells treated with PC 15% or MTA or control cells (Figure 6).
transcription factor mRNA increased by 12-18-fold in differentiated OLC and EXP-21 cells with respect to the undifferentiated hDPSC and remained high at all of the evaluated points, whereas the OSX transcripts reached a significant peak after 7 days of OLC differentiation (200-fold change).
Figure 7(b) shows the results of RUNX2
and OCN assays.
The result showed that, in both arrangements, without use of osteogenic medium, there was an increase in bone related genes RUNX2
Meng et al., "MiR-30a inhibits osteolysis by targeting RunX2
in giant cell tumor of bone," Biochemical and Biophysical Research Communications, vol.
To further characterize the phenotype of POLCs, we investigate the expression of the main markers of osteoblasts, RUNX2
, RANKL, and VDR.
One such example is maternal expression gene 3 (MEG3) regulating the expression of Bmp4, Runx2
, and Osx in human mesenchymal stem cells [12, 13].
The membranes were then incubated with primary antibodies (Runx2
, PPAR[gamma]-2, GSK-3[beta], [beta]-catenin, Dkk-1, and GAPDH, all 1: 1000 dilution) overnight at 4[degrees]C and washed 3 times with TBST for 5 min each before incubation with the secondary antibody at room temperature for 1 h.
After osteogenic differentiation was induced in MSCs cultured for 21 days, the expression of osteogenesis-associated proteins RUNX2
, OPN, and OCN, was analyzed with Western blotting.