Novel functional roles of uncharacterized genes as functional regulators of cellular cholesterol levels. Novel functional roles of uncharacterized genes.

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Volume 19, Issue 5, Pages (May 2017)
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Novel functional roles of uncharacterized genes as functional regulators of cellular cholesterol levels. Novel functional roles of uncharacterized genes as functional regulators of cellular cholesterol levels. (A) Heatmap of drug‐induced gene expression changes (fold change) of CODIM2. Black boxes indicate gene membership of cell‐line‐specific modules. Hierarchical clustering of genes from CODIM2 shows two major components (labeled in blue and green) and three smaller groups (dark green, brown, light blue). (B) Projection onto the STRING protein–protein network reveals a densely interconnected component (enriched in blue set) corresponding to fatty acid and (chole‐)sterol metabolism (Supplementary Table 5 and Supplementary Figure 7). The second component (enriched in green set) is enriched in ER‐related processes, such as protein N‐linked glycosylation, transport and unfolded protein response. Genes subjected to functional knockdown assays of LDL uptake and free cellular cholesterol levels are highlighted in red or orange depending on whether validated as a functional regulator or not, respectively (two or more siRNAs having consistent effect with absolute z‐score >1 and FDR‐corrected P‐value <0.01, Supplementary Table 6). (C, D) Representative image of fixed cells taken by automated widefield fluorescence microscopy. Cells have been transfected for 48 h with siRNA targeting indicated gene, and experiments of (C) LDL‐uptake assay with fluorescently labeled DiI–LDL and (D) staining with cholesterol binding dye filipin was performed. Total intensity of fluorescent signal in segmented spots/area has been quantified as indicated with arrows. Murat Iskar et al. Mol Syst Biol 2013;9:662 © as stated in the article, figure or figure legend