Figure 4. Osteoblast differentiation is sensitive to methionine concentration.

(A) Experimental design. (B) Alizarin red S staining (scale bar = 130 μm). Cells were treated with no supplemental methionine or methionine excess (100 μM and 500 μM) when differentiation initiated, i.e., after transferring the cells to 39.5°C. (C) Alizarin red S staining quantification. Significance assessed relative to control; **P < 0.01, 1-way ANOVA/Dunnett’s multiple-comparison correction (n = 4 biological replicates).