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. 2023 Feb 8;8(3):e165369. doi: 10.1172/jci.insight.165369

Figure 4. Hepatocyte Notch activity regulates MCP-1.

Figure 4

(A) Ccl2 expression in Notch-inactive and -active hepatocytes isolated from NASH diet–fed transgenic Notch reporter male mice (n = 8 mice/group). (B) Comparative sequence alignment of MCP-1 promoter, with evolutionarily conserved Rbpj binding site indicated. (C) Experimental schematic for chromatin immunoprecipitation (ChIP) experiment, showing primer pairs that include (F2/R2) or bind outside (F1/R1 and F3/R3) the Rbpj binding site in the MCP-1 promoter. (D and E) Rbpj occupancy at the MCP-1 promoter in livers from chow- and NASH diet–fed WT mice (n =3 mice/group). (F) Rbpj occupancy at the MCP-1 promoter in livers from Cre- control and hepatocyte-specific Notch gain-of-function (L-NICD) male mice (n = 3 mice/group). (G) MCP-1 promoter-luciferase activity from Ad-GFP and Ad-NICD transduction of mouse primary hepatocytes (n = 3 biologic replicates/group). All data are shown with group means ± SEM; **, P < 0.01, ***, P < 0.001, ****, P < 0.0001 by 2-tailed t test.