Перегляд вихідного коду сторінки Was Roxadustat Worth The Dough?

We validated the specificity of the antibody using a number of assays (see Extended Experimental Procedures; Figures S4A�CS4C) and showed that it can also detect the secreted form of the protein in 293T cell cultures ( Figure?S4D). Western blot [http://en.wikipedia.org/wiki/Adenylate_cyclase Adenylate cyclase] assays of protein extracts from the?brain of Df(16)A+/? mice and WT littermates showed the expected increase (25%) in Mirta22 levels in mutant mice ( Figure?4B). A similar magnitude increase of the Mirta22 immunocytochemical signal was observed in Df(16)A+/? cultured neurons, as compared to WT neurons ( Figure?4C). Immunostaining of neuronal cultures showed that Mirta22 is primarily a neuronal protein (Figure?4D, top). At the subcellular level, it is found primarily [http://www.selleckchem.com/Androgen-Receptor.html Androgen Receptor Antagonist] in the soma, where it colocalizes with the Golgi apparatus marker GM130 (Nakamura et?al., 1995). As the neurons mature, it is also found in vesicles and tubular-like clusters within the dendritic shafts (Figure?4D, middle). Mirta22 immunoreactivity was not detected in cultures stained with preimmune serum (Figure?S4E) and was diminished by 64% in Mirta22 shRNA-transfected neurons ( Figures S4F and S4G, lower panel). Immunohistochemical analysis demonstrated that Mirta22 is widely distributed in the brain, where it is localized in neurons ( Figure?4D, bottom). Accumulating evidence suggests that miRNAs may target functionally connected genes, often in a developmental stage-specific manner (Tsang et?al., 2010; Zhang et?al., 2009). Consistent with this notion, functional annotation [http://www.selleckchem.com/products/fg-4592.html Roxadustat concentration] clustering analysis of?2,695 out of 2,708 predicted miR-185 targets (TargetScan Mouse v.5.2) included in the DAVID Mus musculus gene functional annotation database (http://david.abcc.ncifcrf.gov) identified as the top enriched gene cluster (gene count?= 159, Enrichment Score?= 8.56, FDR-corrected p?= 2?�� 10?9) the Gene Ontology (cellular component) term ��Golgi apparatus�� ( Figure?5A). Gene set enrichment analysis (GSEA) on the 2,708 predicted miR-185 targets ranked based on the gene expression profile of Df(16)A+/? mice also indicated that the Gene Ontology terms ��Golgi apparatus part�� and ��Golgi apparatus�� were among the top 20 gene sets in the adult HPC ( Figure?5A). A global perspective on the enrichment of this miR-185 target gene set among the differentially expressed genes in the Df(16)A+/? mice showed a significant enrichment in the adult HPC expression profile (p?= 5?�� 10?4) where, as expected, most of the top genes were upregulated (n?= 34), and only four genes were downregulated (p?