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. 2019 Aug 12;9:11585. doi: 10.1038/s41598-019-47737-w

Figure 7.

Figure 7

Liver spheroids derived from the TM6SF2 E167K donors show substantially decreased synthesis and secretion of APOB as compared to spheroids derived from the WT donors. (A) Immunoblots using anti-APOB and anti-calnexin (CNX) antibodies comparing APOB synthesis (lysate) and secretion (medium) in WT-1, TM6SF2 E167K-2 and WT-2 show decreased APOB particle number intracellularly and in medium samples obtained from the TM6SF2 E167K-2 donor after incubation with high FA (BSA + 213 µM OA/107 µM PA) for up to 10 days. Quantifications relative to WT-1 and WT-2 are shown below the cropped blots. Original, uncropped blots are provided in Supplementary Fig. S9A. (B) Immunoblot and quantifications layout as presented in (A), showing donor WT-3 vs. TM6SF2 E167K-1. Original, uncropped Blots are provided in Supplementary Fig. S9B. (C) Relative mRNA levels of APOB and MTTP (log2 transformed) after a 2-day incubation with high FA (BSA + 213 µM OA/107 µM PA), normalized to GAPDH and expressed relative to the levels of FA-free BSA treated control. APOB and MTTP are slightly downregulated in spheroids derived from the TM6SF2 E167K donors, while the WT-1 and WT-2 donors show a stronger downregulation of APOB and MTTP. In WT-3, APOB expression was unchanged and MTTP slightly downregulated. The figure represents data from one experiment per donor and RNA was extracted from 20 pooled spheroids per donor. Abbreviations: Ab’, antibody; APOB, APOLIPOPROTEINB100; CNX, calnexin; FA, fatty acid; MTTP, microsomal triglyceride transfer protein; OA, oleic acid; PA, palmitic acid.