Figure 5

Quantitative real-time PCR. Total RNA was extracted from TSA and BMP2 treated neurosphere cultures. Neurosphere cultures were cultured for 7 days and subsequently dissociated and plated out in monolayers. Cells were treated with TSA (10, 25, 50nM) or 10 ng/μl BMP2 from 1.5-2.5 days after plating. Total RNA was collected 6, 12 or 24 h upon treatment. Reverse-transcript cDNA was analyzed using primer recognizing the following genes: Bmp2 (A), Bmp4 (B), Id1 (C), Id2 (D), Smad7 (E), Stat3 (F). Gpr17 (G), Bambi (H), Wnt5a (I), Wisp1 (J) and Primer recognizing beta-Actin and Gapdh were used for normalization. Shown values represent the mean (+/− SEM) of three individual experiments (6 h and 24 h) or two individual experiments (12 h). * = p < 0.05, ** = p < 0.01, *** = p < 0.001, Student’s T-test.