Supplementary Figure 6: IRE1α regulates MAMs biology in vivo.
(A) Upper panel: scheme representing Ern1 structure and deletion strategy (Ern1ΔR). Down, left: livers from Ern1 and Ern1ΔR processed for WB for the indicated proteins. Western blot (WB) is representative of a minimum of three independent experiments. Bottom panel, right: Ern1 and Ern1ΔR mice were intraperitoneally injected with 1 mg/Kg tunicamycin (Tm) or vehicle for 6 h and then Xbp1 mRNA splicing was evaluated by RT-PCR (n = 2 independent experiments). (B) Illustration of the flow chart of the MAMs proteomics. Left: representative WB of wild-type liver subcellular fractionation highlighting the MAMs fraction. Middle: silver staining of representative MAMs fractions. Right: Venn diagram representing the intersections between the hits obtained in the MAMs proteomics of this study, with two other studies, (related to Supplementary Tables 3 and 4). (C-D) Indicated liver samples were processed to obtain subcellular fractions and analysed by WB against the indicated antibodies. (C) Total extracts (Ern1 n = 6 animals; Ern1ΔK n = 5 animals). (D) ER fraction (Ern1 n = 5 animals; Ern1ΔK n = 4 animals). (E) Indicated liver samples were processed to obtain subcellular fractions and analysed by WB for the indicated markers (Cr: crude mitochondria; H: homogenate; M: MAMs; P: pure mitochondria; C: cytosol; E: ER) Representative WB of n = 3 animals. All plots represent mean and SEM. Source data have been provided in Supplementary Tables 3 and 4.
