Expression of GNAS transcripts and key osteogenic markers during induction of osteogenesis in vitro
(A–F) MSC-like cells were incubated in DMEM or OM and total RNA was extracted to measure (A) GNAS, (B) ALPL, (C) Sp7, (D) BGLAP, (E) IBSP, and (F) TBP mRNA at the indicated time points. Bar graphs represent expression of the target mRNA relative to the HPRT housekeeping gene with or without OM. mRNA values on day 0 are considered to be 100%. Each data point represents the arithmetic mean ± SD of three technical replicates. Data from one representative experiment of three independent experiments are shown. (G) Alkaline activity was visualized in MSC-like cells following incubation in DMEM or OM in triplicate on 12-well plates. Data are representative of three images captured with a 10× objective. Bar graphs depict quantification of images showing ALP activity of MSC-like cells following incubation with DMEM or OM medium. Data are normalized to DMEM incubation medium and each point represents the arithmetic mean ± SD of three technical replicates. Data from one representative experiment of three independent experiments are shown (∗p < 0.05). Images only from MEFs cells are shown. OM, osteogenic medium.