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. Author manuscript; available in PMC: 2018 May 1.
Published in final edited form as: Bone. 2017 Mar 8;98:38–46. doi: 10.1016/j.bone.2017.03.003

Figure 2.

Figure 2

A: Proliferation rate of WT and ank/ank BMSCs cultured for 12 h, 24 h and 48 h using the Cell counting Kit-8 (CCK-8). After the incubation with CKK-8 solution, the absorbance of the collected medium was measured at 450 nm at various time points. All the absorbance rates are expressed as percent of the absorbance rate of WT cells after 12 h culture period, which was set as 100%. Data were obtained from 3 different cultures and are expressed as mean ± S.D. (ap < 0.01 vs. WT cells cultured for 24 h; bp < 0.01 vs. WT cells cultured for 48 h). B: ALP activity and alizarin red S staining of WT and ank/ank BMSCs cultured for 7, 14 and 21 days in growth medium (GM) or osteogenic differentiation medium (ODM). C: mRNA levels of ALP (early osteogenic marker), osteocalcin (OC, late osteogenic marker) and the two osteogenic transcription factors osterix and runx2 in WT and ank/ank BMSCs cultured for 1d, 3d, 7d and 14d in GM or ODM. mRNA levels were determined by real time PCR using SYBR Green and normalized to the level of 18S RNA. The mRNA levels of ALP, osterix and runx2 are expressed as relative to the levels of WT BMSCs cultured in growth medium for 1 day, which were set as 1. mRNA levels of OC are expressed as relative to the levels of WT BMSCs cultured in ODM for 7 days, which was set as 1. mRNA levels of OC were not detectable (n.d.) in WT BMSCs cultured in GM, WT BMSCs cultured in ODM for 1 and 3 days, and ank/ank BMSCs cultured in GM or ODM. Data were obtained from triplicate PCRs using RNA from 3 different cultures. Data are expressed as mean ± S.D. (ap < 0.01 vs. WT cells cultured in GM; bp < 0.01 vs. WT cells cultured in ODM).