Skip to main content
. 2019 Jan 4;116(4):1279–1288. doi: 10.1073/pnas.1812588116

Fig. 3.

Fig. 3.

While ZDHHC5 does not affect processing of other Furin/PC7 substrates, it does affect cleavage in endosomes and lipid rafts. (A) HA-tagged E-cadherin was transfected into RPE-1 cells that were silenced for Control, Furin (F here), PC7, F/PC7, or ZDHHC5. Western blots using HA antibody show both uncleaved (u) and cleaved (c) versions of E-cadherin. Statistics: unpaired two-tailed t test. (B) Representative blot of IGF-1Rβ levels in cells silenced for F/PC7 or ZDHHC5, with or without transfection of Furin-V5. Both uncleaved (u) and cleaved (c) IGF-1Rβ are labeled. (C) The FLAG-tagged late endosome (LE-FLAG) sensor was transfected into RPE-1 cells and the PC-specific inhibitor chloromethyl ketone (CMK) was added for 6 h at 50 μM, after which cells were lysed and subjected to SDS/PAGE, transferred, and probed with an anti-FLAG antibody. (D) PC FLAG-tagged sensors targeting different subcellular compartments (PM for plasma membrane, TGN for trans-Golgi network, and GPI for glycosylphosphatidylinositol-anchored proteins primarily found in lipid rafts) were transfected in cells either silenced for Control or ZDHHC5. Cells were lysed and the levels of the cleaved sensors were measured using Western blots with the anti-FLAG antibody. Statistics: unpaired two-tailed t test. *P < 0.05, **P < 0.01.