Bone Sialoprotein Antibody - #DF7738

FIGURE 1Metformin activates AMPK and calcification inVSMCs. VSMCs were cultured with high phosphate (3 mM Pi) and metformin (met)for 7 days. (a) Calcium content (mg/mg protein) of cells treated with metformin (0.5−1.5 mM), (n= 3). (b) Representative alizarin red images ofcells treated with metformin (1 mM). (c) Representative immunoblots (d) immunoblot quantification showing the effect of metformin (1 mM) onexpression of osteogenic markers osterix, osteocalcin, and bone sialoprotein compared withβ‐actin (n= 3). (e) Representative immunoblots and(f) immunoblot quantification showing the effect of metformin (1 mM) on the phosphorylation of AMPK compared with total AMPK (n= 4).AMPK, adenosine monophosphate‐activated protein kinase; VSMCs, vascular smooth muscle cells.PHADWALET AL.|4307

Figure 1 Metformin inhibits calcification in RVICs and reduces the induction of RUNX-2 and bone sialoprotein expression. (A) Calcium content (mg/mg protein) of cells treated with metformin (Met) (0.5–1.0 mM), (n = 3). (B) Alizarin red staining and its quantification in cells treated with metformin (1 mM), (n = 3). (C) mRNA expression of Runx2 in calcified RVICs cultured in presence/absence of metformin (1 mM) for 48 and 72 h. (n = 3). (D) Representative western blots and (E, F) western blot quantification showing the effect of metformin treatment (1 mM) on the protein levels of Runx2 and bone sialoprotein compared to β-actin (n = 4). Data shown as mean + / − S.E.M *P 

Figure 1. P. gingivalis obstructs the osteogenic/cementogenic differentiation of PDLCs. PDLCs were cultured with Pg at a MOI of 100 for 7 days, mRNA expressions of OCN, OSX, CAP, CEMP-1 were measured by RT-qPCR (a), and protein expressions of OCN, BSP, OSX, RUNX2 in PDLCs were detected by western blotting (b). PDLCs were incubated with Pg at a MOI of 200 for 7 or 14 days, and ALP staining on day 7 (c), SRS on day 7 (e) and ARS (g) on day 14 were conducted and quantified (d, f, h). Pg: P. gingivalis; PDLCs: periodontal ligament cells; MOI: multiplicity of infection; OCN, osteocalcin; BSP, sialoprotein; CAP, cementum attachment protein; CEMP-1, cementum protein-1; OSX: osterix; RUNX2: runt-related transcription factor 2; RT-qPCR:: reverse transcription quantitative polymerase chain reaction; ALP: alkaline phosphatase; SRS: sirius red staining; ARS: alizarin red staining. Significance was defined as *P 

Fig. 4. SMF promoted the osteogenic differentiation of PDLSCs. a mRNA expression of ALP, BSP, and RUNX2was detected by qRT–PCR after cells were cultured for 7 days. b, c The protein expression levels of ALP, BSP, and RUNX2 were detected by Western blot after cells were cultured for 7 and 14 days. All experiments were performed in triplicate, and the values are expressed as the mean ± SD. *p< 0.05, **p< 0.01.