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. 2014 Sep 4;9(9):e106661. doi: 10.1371/journal.pone.0106661

Figure 3. Whsc1 deficiency inhibits osteoblast differentiation but not cell proliferation.

Figure 3

(A, B) Alp (A) and Ocn (B) mRNA expression during bone differentiation was measured by qRT-PCR. MC3T3-E1 cells were transfected with the indicated siRNAs and differentiated by incubation with BMP-2. Values were normalized to the expression of 18S. (C, D) Alp activity was assessed using an alkaline phosphatase assay (C) or through Alp staining (D) in MC3T3-E1 cells transfected with the indicated siRNAs and in the presence or absence of BMP-2. (E) mRNA expression during bone differentiation was measured by qRT-PCR. MC3T3-E1 cells transfected with the indicated siRNAs were differentiated by 3 days (BMP3) and 6 days (BMP6) incubation with BMP-2. Values were normalized to the expression of 18S. (F) MC3T3-E1 cells transfected with the indicated siRNAs were analyzed by western blot analysis with the indicated antibodies. (G) Cell proliferation of MC3T3-E1 cells transfected with the indicated siRNAs. MC3T3-E1 cells were differentiated by 6 days incubation with BMP-2. The value of wild type was set to 1.0. For all panels, Error bars represent the s.d. *, P<0.05. **, P<0.01.