Figure 4. P19T1A2 cells express Ascl1 and differentiate in response to Dox in a dose-dependent manner.
A. Ascl1 (red) staining of P19T1A2 cells treated with the indicated concentrations of Dox for 24 h. Treatment of P19T1A2 cells with as low as 3 ng/ml of Dox resulted in sporadic Ascl1-positive cells, and the number and intensity of Ascl1-positive cells increased with greater concentrations of Dox. B. TuJ1 (red) images P19T1A2 cells treated with the indicated concentrations of Dox. P19T1A2 cells expressed TuJ1 in a dose-dependent manner. Furthermore, in the highest concentrations of Dox, the total number of cells was reduced due to the inhibition of proliferation resulting from neuronal differentiation. Scale bars = 100 μm. C, D. Western blot analysis for expression of Ascl1 (C) and β-III-tubulin (D) protein showed that protein expression correlates with the results shown in (A and B).