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. 2016 Apr 15;27(8):1220–1234. doi: 10.1091/mbc.E15-08-0557

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© 2016 DeRossi, Vacaru, et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 5: — Terpenoid and dolichol synthesis defects in trappc11 mutants. (A) Schematic representation of the terpenoid biosynthetic pathway, with branch points at farnesyl-PP leading to production of dolichol (protein glycosylation), squalene (steroid synthesis), and geranylgeranyl-PP (protein prenylation). Fold changes of expression in trappc11 mutant livers are indicated by color: blue indicates decrease and red indicates increase relative to WT siblings; gray indicates genes not tested. Drugs used are indicated with a red T at their targets. (B) Confocal microscopy of transgenic WT and trappc11-mutant zebrafish expressing a prenylated form of GFP (Tg(actb2:CAAX-GFP)) in hepatocytes. Zebrafish were collected at 5 dpf and analyzed for membrane localization of CAAX-GFP. Samples were counterstained with DAPI (nucleus) and Cy5-streptavidin to stain the biotin-rich hepatocytes (middle; white boxes outline magnified GFP-only images shown on right). Scale bars, 10 μm. (C) Confocal microscopy of WT transgenic zebrafish expressing a prenylated form of GFP in hepatocytes (Tg(fabp10:CAAX-GFP). Zebrafish larvae were treated with DMSO (top) or 0.75 μg/ml BFA (bottom) from 3 to 5 dpf and analyzed at 5 dpf for membrane localization of CAAX-GFP. Samples were counterstained with DAPI (blue nucleus). White boxes outline area shown magnified on right (GFP only). Scale bars, 10 μM. (D) Whole WT and trappc11-mutant larvae were collected at 5 dpf, and total cholesterol was measured and normalized to protein content. trappc11 mutants have a significant decrease in cholesterol compared with WT siblings (p = 0.0008, paired Student’s t test), suggestive of decreased terpenoid biosynthetic activity. Individual clutches are indicated (n = 17 for each), with lines representing clutch pairing.