The authors of “Angiopoietin-like 4 (ANGPTL4) promotes intracellular degradation of lipoprotein lipase in adipocytes” (J. Lipid Res. 2016. 57: 1670–1683) have advised the Journal that the last sentence of each of the figure legends for Figs. 5, 6, 7, 8, 9, and 10 is incorrect. The descriptions within parentheses of the R and S gel bands were inadvertently swapped. The figure legends for these figures should read as follows:
Fig. 5.
ANGPTL4 lowers the amount of LPL on the adipocyte cell surface. A: Western blot of cell lysates of gWAT explants from Angptl4−/− and wild-type mice incubated in the absence or presence of heparin (50 IU/ml) for 3 h. Western blots were probed with antibodies against LPL and HSP90 (as loading control). B: Western blot of cell lysates of adipocytes that had been differentiated from stromal vascular fractions of WAT from Angptl4−/− and wild-type mice and incubated in the absence or presence of heparin (10 IU/ml) for 20 min. Western blots were probed with antibodies against LPL and HSP90 (as loading control). C: Angptl4 and Lpl mRNA levels in gWAT explants from Angptl4−/− and wild-type mice. D: Angptl4 and Lpl mRNA levels in adipocytes that had been differentiated from stromal vascular fractions of WAT from Angptl4−/− and wild-type mice. 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.
Fig. 6.
ANGPTL4 lowers LPL secretion. A: Western blot of media of gWAT explants from Angptl4−/− and wild-type mice treated in the absence or presence of heparin (50 IU/ml) for 3 h. Western blots were probed with antibodies against LPL and ADIPOQ. Coomassie blue staining was used to assess loading. B: Western blot of media of adipocytes that had been differentiated from stromal vascular fractions of WAT from Angptl4−/− and wild-type mice and incubated in the absence or presence of heparin (Hep.) (10 IU/ml) for 20 min. Western blots were probed with antibodies against LPL and ADIPOQ. Coomassie blue staining was used to assess loading. C: Western blot of media of adipocytes that had been differentiated from stromal vascular fractions of WAT from Angptl4−/− and wild-type mice. Cells were pretreated for 20 min with heparin (10 IU/ml), washed with PBS, and incubated with heparin (10 IU/ml) for the indicated times. Western blots were probed with antibodies against LPL and ADIPOQ. 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.
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.
Fig. 8.
Physiological regulation of ANGPTL4 expression affects levels of EndoH-resistant LPL in vivo. A: Western blot of gWAT lysates of Angptl4−/− and wild-type mice, as analyzed by SDS-PAGE and native PAGE. Western blots were probed with antibodies against LPL and HSP90. Coomassie blue staining was used to assess loading. B: Western blot of EndoH-treated and PNGase-treated gWAT lysates of Angptl4−/− and wild-type mice. Western blots were probed with antibodies against LPL and HSP90 (as a loading control). C: Western blot of EndoH-treated gWAT lysates prepared from ad libitum-fed, overnight-fasted, or refed Angptl4−/− and wild-type mice. The tissue samples were taken from an experiment described earlier (13). Western blots were probed with an antibody against LPL; Coomassie blue staining was used to assess loading. D: Western blot of gWAT lysates prepared from ad libitum fed, overnight-fasted, or refed wild-type mice. The tissue samples were taken from an experiment described earlier (13). Western blots were probed with an antibody against ANGPTL4; Coomassie blue staining was used to assess loading. E: Western blot of EndoH-treated gWAT lysates from Angptl4−/− and wild-type mice that were either not perfused or perfused with PBS containing heparin (50 IU/ml). Western blots were probed with antibodies against LPL and HSP90 (as a loading control). F: Western blot of EndoH-treated gWAT lysates prepared from fed and overnight-fasted Angptl4−/− and wild-type mice that had been given an intravenous injection of heparin (100 IU/kg). 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.
Fig. 9.
Levels of EndoH-resistant LPL are inversely related to Angptl4 expression. A: Western blot on 0.75 μl of plasma from Angptl4−/− and wild-type mice. Western blots were probed with an antibody against LPL; Coomassie blue staining was used to assess loading. B: Western blot of EndoH-treated WAT lysates prepared from Angptl4−/−, Angptl4+/-, and wild-type mice. Western blots were probed with an antibody against LPL; Coomassie blue staining was used to assess loading. C: Western blot of EndoH-treated WAT lysates prepared from Angptl4−/−, wild-type, and Angptl4-Tg mice. Western blots were probed with an antibody against LPL; Coomassie blue staining was used to assess loading. D: Western blot of EndoH-treated lysates of 3T3-F442a adipocytes that had been treated with rosiglitazone (10 μM) or DMSO control for 6 h. Western blots were probed with antibodies against LPL, ANGPTL4, and HSP90 (as a loading control). E: Western blot of EndoH-treated lysates of 3T3-L1 adipocytes that had been treated with rosiglitazone (10 μM) or DMSO control for 6 h. Western blots were probed with antibodies against LPL, ANGPTL4, 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.
Fig. 10.
ANGPTL4 regulates levels of EndoH-resistant LPL in adipose tissue, but not heart. A: Western blot of EndoH-treated BAT lysates from Angptl4−/− and wild-type mice exposed to cold (4°C) or thermoneutral temperature (28°C) for 10 days. The tissue samples were taken from an experiment described earlier (14). Western blots were probed with antibodies against LPL and HSP90 (as a loading control). B: Western blot on EndoH-treated heart lysates from Angptl4−/− and wild-type mice fed ad libitum or fasted overnight. 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.