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. 2021 Dec 16;13(3):879–899. doi: 10.1016/j.jcmgh.2021.12.008

Figure 7.

Figure 7

Localization of TM6SF2 to the ER and ERGIC using cell fractionation. (A) A diagram of the cell fractionation protocol (left) and immunoblot of fractionated liver lysates (right). Rat liver homogenate (0.5 g) was loaded on a discontinuous sucrose gradient and centrifuged at 100,000g for 1 hour, yielding a light fraction (Golgi) and heavy fraction (ER). Equal proportions (0.8% of total volume) of each fraction were subjected to immunoblot analysis. (B) A diagrammatic scheme of the cell fractionation protocol (left) and immunoblot analysis of cell fractionation of liver lysates (right). Golgi membranes were isolated from livers of WT and Tm6sf2 KO rats after a 16-hour fast. A proportion of each fraction (as indicated) was subjected to immunoblot analysis. All experiments were repeated at least twice and results were similar. ∗Nonspecific band. BIP, binding immunoglobulin protein; CANX, calnexin; COX IV, cytochrome oxidase IV; EEA1, early endosome antigen 1; G, Golgi-enriched fraction; GOS28, Golgi SNAP receptor complex member 1; H, homogenate; I, intermediate fraction; LAMP1, lysosomal-associated membrane protein 1; LDH, lactate dehydrogenase A chain; LSD1, lysine-specific demethylase-1A; Plin2, Perilipin 2; PMP70, 70-kilodalton peroxisomal membrane protein.