(A) Osteocalcin (OCN) expression in mature MM-derived OBs, with or without CCL3 (50 ng/ml), detected via (left panel) quantitative PCR analysis, (middle panel) ELISA on culture supernatant, and (right panel) immunofluorescent detection of OB cultures (OCN is stained with Alexa Fluo 488, counterstain with DAPI). (B) Western blot detection of pERK, pAKT, pP38, ERK1/2 on HS27A-derived OB exposed to CCL3 50 ng/ml for the specified time points. (C) Western blot detection of pERK and ERK1/2 in HS27A-derived OB after dose-dependent exposure to CCL3. A densitometric analysis averaging more than 3 experiments is shown. (D) Osterix gene expression assessed by quantitative PCR in OB exposed to increasing time and doses of CCL3. (E)
RUNX2 and ATF4 gene expression evaluation on mature OBs in the presence of CCL3 (4h, 50 ng/ml) (* p<0.05, ** p<0.01)