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. 2019 Nov 13;4(1):77–91. doi: 10.1002/hep4.1450

Figure 4.

Figure 4

HSCs carrying the PNPLA3 I148M SNP induce a more pro‐inflammatory and steatotic phenotype in the MPS NASH model. PHH, HK, and HSC co‐cultures were cultured in the MPS for 15 days in HEP‐LEAN, HEP‐FAT, or HEP‐FAT + LPS conditions (LPS was added to culture from day 8 onward). Cultures contained either WT HSC or HSC with the PNPLA3 I148M mutation. (A) On day 8 (pre‐LPS dose) and day 15 (post‐LPS dose) of the culture, IL‐6 production was measured by ELISA. On day 15 of the culture, albumin production was measured by ELISA (B), and fat loading was measured by Oil Red O staining of microtissues (C), which was quantified by absorbance at 510 nm to give a relative fat content. (D) Relative expression of total RNA, as measured by quantitative PCR; expression of each gene was normalized to GAPDH and is shown as relative to lean WT samples. (E) Procollagen 1 and TIMP‐1 production was measured by ELISA. Data are presented as means ± SD for 12 independent cultures (3 HSC donors per condition and n = 4 per donor); *P < 0.05. Abbreviations: ACTA2, smooth muscle alpha‐2; GK, Glycerol Kinase; IGFBP1, insulin‐like growth factor‐binding protein 1.