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RNAi–Based Functional Profiling of Loci from Blood Lipid Genome-Wide Association Studies Identifies Genes with Cholesterol-Regulatory Function

Figure 4

Impact on FC levels and subcellular localization of GFP–tagged candidate genes.

(A) cDNAs encoding for indicated candidate genes linked to GFP were transiently expressed in Hela-Kyoto cells and impact on cellular FC levels was analyzed (see Materials and Methods, Figure S6 and Table S8 for comprehensive datasets). Arrows denote “transfected”, arrowheads “untransfected” cells. See Materials and Methods for definition of threshholds (dashed lines in graphs).Graphs depict total segmental filipin signal plotted against total cellular intensities in the GFP-channel. Each dot reflects one individual cell, trend lines are given in red. Numbers indicate mean ratios of FC in GFP-positive relative to non-expressing cells within the identical dish (n = 3–4 experiments). (B) Maximal projections of confocal stacks showing representative GFP-cDNA expressing cells under control and sterol-depleted conditions (see Materials and Methods). Arrows denote cellular compartments with increased signals upon sterol-depletion. Bars = 10 µm.

Figure 4