Fig. 7.

ANGPTL4-mediated loss of LPL protein occurs in a postER compartment. A: Western blot of EndoH-treated cell lysates of adipocytes that had been differentiated from stromal vascular fractions of WAT from Angptl4^−/− and wild-type mice. Cells were treated with 5 μg/ml brefeldin A for 2 h. Western blots were probed with antibodies against LPL and HSP90 (as a loading control). B: Western blot of cell lysates of adipocytes that had been differentiated from stromal vascular fractions from WAT of Angptl4^−/− and wild-type mice. Cells were lysed in NP-40 lysis buffer. Western blots were probed with antibodies against LPL, HSP90 (as a loading control), and H2A (as a loading control). NP40S, NP40-soluble LPL; NP40P, NP40-precipitated LPL; s.e., short exposure; l.e., long exposure. C: Western blot of EndoH-treated cell lysates of adipocytes that had been differentiated from the stromal vascular fractions of WAT from Angptl4^−/− and wild-type mice. Cells were treated with 10 μM monensin (Mon.) for 3 h. Western blots were probed with antibodies against LPL and HSP90 (as a loading control). D: Western blot of EndoH-treated cell lysates from adipocytes that had been differentiated from stromal vascular fractions of WAT from Angptl4^−/− and wild-type mice. The cells were treated with 20 μM E64D for 24 h. Western blots were probed with antibodies against LPL and HSP90 (as a loading control). E: Western blot of EndoH-treated cell lysates of adipocytes that had been differentiated from stromal vascular fractions of WAT from Angptl4^−/− and wild-type mice. Cells were treated with 5 mM 3MA for 10 h. Western blots were probed with antibodies against LPL and HSP90 (as a loading control). F: Western blot of EndoH-treated cell lysates of adipocytes that had been differentiated from stromal vascular fractions from WAT of Angptl4^−/− and wild-type mice. The cells had been treated with 10 mM leupeptin (Leup.) for 10 h. Western blots were probed with antibodies against LPL and HSP90 (as a loading control). EndoH-resistant LPL (complex oligosaccharides; Golgi and cell surface LPL) is indicated with R; EndoH-sensitive LPL (high-mannose oligosaccharides, ER LPL) is indicated with S.