GKS proteins are enriched on LDs in IRGM-deficient cells (original) (raw)

2013

Abstract

<p>(<b>A</b>) Wildtype MEFs were treated overnight with OA and IFNγ and immunostained for endogenous Irgm3 and for LDs using BODIPY. (<b>B</b>) Wildtype MEFs were transfected with the three mouse paralogs Irgm1-3 tagged with V5, infected with <i>C. trachomatis</i> and treated with OA and IFNγ at 3 hpi. Cells were fixed at 20 hpi and stained with anti-V5, BODIPY and Hoechst. White arrows point at inclusions. (<b>C</b>) The localization of the GKS proteins Irga6 and Irgb6 were visualized in wildtype and <i>Irgm1/m3^−/−</i> MEFs treated overnight with OA and IFNγ. (<b>D</b>) Cells were treated overnight with OA and IFNγ and stained with anti-Irgb10, BODIPY and Hoechst. Representative images are shown. Overlap in fluorescence signals derived from anti-Irgb10 and BODIPY stains is shown for these representative images. Data were analyzed as described in <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1003414#s4" target="_blank">Materials and Methods</a> and representative data of three independent experiments are shown. Statistical significance of group values relative to wildtype and between marked groups is shown (***, p<0.005). (<b>E</b>) LDs and total cell lysate were obtained from wildtype, <i>Irgm1/m3^−/−</i> and <i>Atg5</i>^−/− MEFs treated overnight with OA +/− IFNγ and analyzed by immunoblotting.</p

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