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. 2018 Sep 12;2018:2190657. doi: 10.1155/2018/2190657

Figure 3.

Figure 3

Osteogenic differentiation of hASCs in BM, OM, and AM culture conditions supplemented with FAK, ERK, and ROCK inhibitors. (a) The cells were cultured in BM, OM, or AM supplemented with 2 μM FAK, 40 μM ERK, or 15 μM ROCK inhibitors in addition to medium controls. RUNX2A expression was analyzed with qRT-PCR at 7 d. FAK and ROCK: N = 5 (independent experiments, 5 donors), ERK: N = 3 (independent experiments, 3 donors). (b) ALP activity was analyzed with ALP assay at 7 d and 14 d. The ALP absorbance values were normalized with corresponding CyQUANT results, and the results are presented relative to the 7 d BM sample. Significance level 5%, designated with an asterisk (∗). FAK, ERK, ROCK: N = 9 (independent biological replicates from 3 donors). (c) Matrix mineralization was analyzed with AR staining after 14 d and 21 d of culture. Quantitative results of AR staining are presented as graphs and corresponding representative images of the stained wells (21 d, area 1.9 cm2) are presented below; bright red dye represents mineral. Significance level 5%. FAK, ROCK: N = 18 (independent biological replicates from 6 donors, control condition values of the graphs are the same since the experiments were conducted at the same time), ERK: N = 15 (independent biological replicates from 5 donors). BM: basic medium; OM: osteogenic medium; AM: adipogenic medium; ALP: alkaline phosphatase; AR: Alizarin Red.