Ation developmental process blood vessel development TGF beta signaling pathway organ
Ation developmental process blood vessel development TGF beta signaling pathway organ development cell fate determination mesoderm development negative regulation of developmental process0 1,0 2,0 3,0 4,0 5,0 6,0 7,0 8,0 9,enrichment scoreenrichment scoreFigure 4 Functional annotation order TAK-385 clustering of BMP2 and TSA treated neurosphere cultures at different time points. Differentially expressed genes (relative expression >2-fold and absolute difference >100. comparing treated and control sample, PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/25679764 confidence >90 ) are functionally clustered using DAVID Database. Functional groups with an enrichment score >1.5 are exhibited. The enrichment score is the geometric mean (in -log scale) of member’s p-values in a corresponding annotation cluster. The number of genes in each cluster is represented as bars, the corresponding enrichment scores are illustrated as dots. Functional annotation of genes altered in individual treatments with TSA after 6 h (A) and 24 h (B), BMP2 after 6 h (C) and 24 h (D) as well as of intersectional genes regulated after treatment with BMP2 24 h and TSA 24 h (E) are displayed.the sandwich ELISA microarray disclosed a clear regulation of Erk2 phosphorylation upon both BMP2 and TSA treatment (Figure 6C). At the 6 h and 12 h time point pErk2 was induced in a concentration dependent mannerafter TSA treatment, but also after BMP2 treatment. After 24 h of treatment the pErk2 signal clearly decreased, which suggested that pErk2 is involved in the early signaling following BMP2 and TSA treatment.Scholl et al. BMC Genomics 2012, 13:298 http://www.biomedcentral.com/1471-2164/13/Page 11 ofA 4,5 Bmp4,0 3,5 3,0 2,5 2,0 1,5 1,0 0,5 0,B**1,2 1,0 0,8 0,6 0,4 0,BmpC 5,0 Id4,5 4,0 3,5 3,* *** ** ***** *** *2,5 2,0 1,5 1,0 0,5 0,control10nM25nM50nM10ng/ml BMP0,control10nM25nM50nM10ng/ml BMPcontrol10nM25nM50nM10ng/ml BMPTSATSATSAD 3,0 Id2,5 2,0 1,5 1,0 0,5 0,E 16,0 Smad* * ** * * *14,0 12,0 10,0 8,0 6,0 4,0 2,F 3,0 Stat2,5 2,* ** ** ** **1,5 1,****control* *****0,**50nM 10ng/ml BMP0,control10nM25nM TSA50nM10ng/ml BMP0,10nM25nM TSAcontrol10nM25nM TSA50nM10ng/ml BMPG 1,8 Gpr1,6 1,4 1,2 1,0 0,8 0,6 0,4 0,2 0,H10,0 Bambi9,0 8,0 7,0 6,0 5,0 4,0 3,0 2,0 1,0 0,IWnt5a14,0 12,0 10,0 8,*** ** ******* *6,0 4,0 2,*control 10nM 25nM TSA 50nM 10ng/ml BMPcontrol10nM25nM TSA50nM10ng/ml BMPcontrol10nM25nM TSA50nM10ng/ml BMP0,J 90,0 Wisp80,0 70,0 60,0 50,0 40,0 30,0 20,0 10,0 0,**6h 12 h 24 h* **control*25nM TSA*50nM*10ng/ml BMP10nMFigure 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/l 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.Discussion We previously demonstrated that treatment of neuronal precursor cells derived from the ganglionic eminences (GE) with BMP2 or TSA resulted in a reduction in the generation of neurons and oligodendrocytes and in an increase in the.
