Figure 5: 7E promoted osteoblast differentiation in hAFSCs.
(a,b) hAFSCs were incubated with osteoblast differentiation medium. The osteoblasts were characterized using ALP staining (a) 14 days post-incubation, or Alizarin staining (b) 28 days post-incubation. (c) Cell lysates were collected and measured; ALP activity was measured using an assay kit 14 days post-incubation. Values are means ± SD. *P < 0.05 versus untreated controls (Ctrl). (d–f) hAFSCs were cultured under osteogenic conditions for 14 days. mRNA was isolated and the transcripts of OSX, RUNX2, and Atf4 were measured using RTQ-PCR with specific primers. The quantification analysis of mRNA was normalized; GAPDH was the housekeeping gene. *P < 0.05 versus untreated controls (Ctrl). #P < 0.05 versus the 7E-treated group. (g) hAFSCs were cultured under osteogenic conditions for 28 days. The expression of sclerostin was analyzed using RTQ-PCR with specific primers. The quantification analysis of mRNA was normalized; GAPDH was the housekeeping gene. *P < 0.05 versus untreated controls. #P < 0.05 versus the IL-20-treated group. All experiments were run three times, with similar results. Data are from a representative experiment.
