FIGURE 3.

Treatment with valproic acid increased the cell size and decreased proliferation and expression of stem cell markers of AM‐1 cells. A‐E, AM‐1 cells were cultured for 24 h with 1 mmol/L valproic acid or 100 mg/ml basic fibroblast growth factor (bFGF). The cells were subjected to ICC using antibodies to β‐catenin (A) and Sox2 (E), and immunoblotting using antibodies to β‐catenin or α‐tubulin (B). Cytoskeletons and nuclei were visualized using Alexa 488‐conjugated phalloidin (green) and DAPI (blue), respectively (A, E). Scale bar = 20 μm. Total RNA of the cells was extracted and subjected to real‐time PCR analyses using primer sets of Bcl11b (C) and Sox2 (D). n = 3. VPA, valproic acid. RQ, relative quantity