Figure 2: (a) Alcian blue staining of micromass. pCAGGS-BMP4 transfected C3H10T1/2 was cultured in micromass for 6 days with LiCl or wnt3a supplementation. (b) Quantitative analysis of Alcian blue staining. LiCl treatment and wnt3a supplementation suppressed chondrogenic differentiation. (c) EGFP expression of pCAGGS-EGFP (30 μg/cuvette) transfected cells at day 3. LiCl did not alter EGFP expression in 5 and 10 mM and stimulated EGFP in 25 mM. (d) Sea pansy luciferase expressing gene, pRL-SV40 (30 μg/cuvette), was transfected. Luciferase activity was measured at day 3. Luciferase activity was not changed in 5 and 10 mM of LiCl treatment and was higher in 25 mM. (e) Cells were cotransfected with TOPFLASH (open bar) or FOPFLASH (solid bar) (20 μg/cuvette), pRL-SV40 (20 μg) and pCAGGS-BMP4 (20 μg). Luciferase activity in cell lysates was measured at day 3. The data was normalized to the activity of cotransfected Renilla control. LiCl treatment dose dependently increased β-catenin in the cells. , , and .