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. 2016 Jun 8;6:27447. doi: 10.1038/srep27447

Figure 4. 5-aza-dC rescues the osteogenic differentiation capacity of hPDLSCs under HG conditions.

Figure 4

(a) hPDLSCs were incubated in osteogenic medium (OM) with 30 mM glucose (HG) or 1 μM 5-aza-dC for 3 or 7 days, and then ALP activity was measured (n = 3). The values (King unit/min/mg protein) are expressed as the mean ± SD. *p < 0.05, **p < 0.01 (Student’s t-test). (b) hPDLSCs were incubated in osteogenic medium (OM) for 2 weeks and mineralization of different groups as indicated was confirmed using alizarin red staining. (c–f) hPDLSCs were incubated in osteogenic medium (OM) with 30 mM glucose (HG) or 1 μM 5-aza-dC for 3 or 7 days, and then mRNA levels of osteogenic markers ALP (c), OCN (d), OPN (e) and OSX (f) were subjected to quantitative RT-PCR analysis (n = 3). GAPDH served as internal control. *p < 0.05, **p < 0.01 (Student’s t-test). (g,h) hPDLSCs were incubated in osteogenic medium (OM) with 30 mM glucose (HG) or 1 μM 5-aza-dC for 3 (g) or 7 (h) days, and then protein levels of ALP, OPN and OSX were measured by Western blot analysis.