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. Author manuscript; available in PMC: 2013 Sep 12.
Published in final edited form as: Leukemia. 2012 Jul 5;27(2):430–440. doi: 10.1038/leu.2012.183

Figure 3.

Figure 3

PIs promote osteogenic differentiation and mineralization in vitro. (a) Primary MSCs from MM patients (6/9 with osteolytic lesions) were cultured in osteogenic medium either in the continuous presence of PIs (left panel) or transiently treated on day 0 for 4 h (right panel). Mineralization was analyzed by alizarin red staining and subsequent dye quantification in OBs differentiated for 21 days; representative images of alizarin red staining following transient treatment are shown. Results are expressed as the mean±s.d. (b) In the presence of PIs and at day 11 of osteogenic differentiation, ALP activity was measured in OBs derived from MSCs from five MM patients (3/5 with osteolytic lesions). Graphs illustrate mean values±s.e.m. (c) Total RNA from the hMSC-TERT cell line was isolated at day 14 of differentiation in the presence of PIs at indicated doses, and expression of osteogenic-related markers (Osterix, osteopontin, osteocalcin) and DKK-1 were evaluated using real-time reverse transcription-PCR. Expression levels for each gene were normalized with respect to GAPDH expression and referred to vehicle control. Data are represented as the mean±s.d. from three different experiments. In all panels: *P<0.05, versus vehicle control or between indicated groups.