antiosteocalcin antibody  (Boster Bio)

Journal: Journal of Agricultural and Food Chemistry

Article Title: Sodium Benzoate Inhibits Osteoblast Differentiation and Accelerates Bone Loss by Regulating the FGF2/p38/RUNX2 Pathway

doi: 10.1021/acs.jafc.5c01124

Figure Lengend Snippet: NaB inhibits MC3T3-E1 cell osteogenic differentiation. (A) ALP staining of MC3T3-E1 cells after induction in OIM containing different NaB concentrations (0, 5, and 10 mM) for 7 days. The right panel shows the quantification of ALP staining, scale bar = 500 μm. (B) ARS staining of MC3T3-E1 cells after treatment with NaB (0, 5, or 10 mM) in OIM for 21 days. The right panel shows the relative OD values of the different groups, scale bar = 500 μm. (C–E) After MC3T3-E1 cells were cultured in OIM for 7 days, images were obtained to show the fluorescence intensity for RUNX2, OCN, and ALP, scale bar = 100 μm. (F) qRT-PCR was performed to evaluate the RUNX2, OCN, and ALP mRNA levels in MC3T3-E1 cells treated with NaB (0, 5, or 10 mM) for 7 days, GAPDH was used as the loading control. (G) The RUNX2, OCN, and ALP protein levels in MC3T3-E1 cells were determined by Western blotting. (H) Quantification and normalization of the indicated protein levels shown in (G); GAPDH was used as the loading control. Data are presented as the mean ± SD * P < 0.05; ** P < 0.01 compared with the 0 mM group.

Article Snippet: Anti-RUNX2 was obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA), antiosteocalcin (OCN), anti-ALP, and anti-α-tubulin antibodies were obtained from Proteintech (Wuhan, China), anti-p38, anti-p-p38, antiextracellular signal regulated kinases 1 and 2 (ERK1/2), anti-p-ERK, anti- c-Jun N-terminal kinases (JNK), anti-p-JNK, and anti-GAPDH antibodies were acquired from Cell Signaling Technology (Danvers, MA, USA), and anti-FGF2 antibody was purchased from Abcam (Cambridge, UK).

Techniques: Staining, Cell Culture, Fluorescence, Quantitative RT-PCR, Control, Western Blot

Journal: Journal of Agricultural and Food Chemistry

Article Title: Sodium Benzoate Inhibits Osteoblast Differentiation and Accelerates Bone Loss by Regulating the FGF2/p38/RUNX2 Pathway

doi: 10.1021/acs.jafc.5c01124

Figure Lengend Snippet: NaB inhibits osteogenic differentiation dependent on p38/MAPK activity. (A) After MC3T3-E1 cells were treated with NaB (10 mM) with or without anisomycin (1 nM) in OIM for 4 days, the p-p38, p38, RUNX2, OCN, and ALP expression levels were determined by Western blotting. (B–E) Quantification and normalization of indicated protein levels shown in (A). (F) ALP staining of MC3T3-E1 cells after treatment with NaB (10 mM) with or without anisomycin (1 nM) in OIM for 7 days. The right panel shows the quantification of ALP staining, scale bar = 500 μm. (G) ARS staining of MC3T3-E1 cells after treatment with NaB (10 mM) with or without anisomycin (1 nM) in OIM for 21 days. The right panel shows the quantification of ALP staining, scale bar = 500 μm. Data are expressed as the mean ± SD ( n = 3). * P < 0.05; ** P < 0.01 compared with the control group.

Article Snippet: Anti-RUNX2 was obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA), antiosteocalcin (OCN), anti-ALP, and anti-α-tubulin antibodies were obtained from Proteintech (Wuhan, China), anti-p38, anti-p-p38, antiextracellular signal regulated kinases 1 and 2 (ERK1/2), anti-p-ERK, anti- c-Jun N-terminal kinases (JNK), anti-p-JNK, and anti-GAPDH antibodies were acquired from Cell Signaling Technology (Danvers, MA, USA), and anti-FGF2 antibody was purchased from Abcam (Cambridge, UK).

Techniques: Activity Assay, Expressing, Western Blot, Staining, Control