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. 2022 Sep 13;25(10):105135. doi: 10.1016/j.isci.2022.105135

Figure 5.

Figure 5

Accumulated palmitic acid in Nudt7+/− liver is responsible for PPARγ activation

(A) The methylation levels of H3K4me2, H3K4me3, H3K27me2, and H3K27me3 were analyzed by immunoblotting in Nudt7+/+ and Nudt7+/− liver. Histone H3 was used for loading control (n = 3).

(B) The transcription levels of Mll3 and Mll4 were analyzed by qRT-PCR (n = 3).

(C) Representative images of H3K4me3 staining in the 12-month-old mouse liver (n = 3; Scale bars, 50 μm) and positive staining ratio (n = 6 per group).

(D) H3K4me3 level on PPARγ promoter was analyzed using qRT-PCR. DNA was isolated using chromatin immunoprecipitation (ChIP) in Nudt7+/+ and Nudt7+/− liver (n = 3).

(E) Palmitic acid level in Nudt7+/+ and Nudt7+/− liver (n = 5).

(F) Primary cultures of Nudt7+/+ hepatocytes were treated with BSA-conjugated with 50 μM palmitic acid (PA) or BSA alone (CON) and the expression level of H3K4me3, H3K27me3, H3K4me2, and H3K27me2 were analyzed by immunoblotting. Histone H3 was used for loading control.

(G) The expression levels of Mll3 and Mll4 were analyzed by qRT-PCR (n = 3).

(H) Primary cultures of Nudt7+/+ and Nudt7+/− hepatocytes were infected with lentiviruses containing NUDT7 (Nudt7-OE, +) or mock control (−) and the expression level of Mll3 was analyzed by qRT-PCR (n = 3).

(I) Representative images of BODIPY493/508 staining in primary cultures of Nudt7+/− hepatocytes transduced with shMll3 for 24 h. Scale bars, 100 μm. ∗p ≤ 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001 (Unpaired t-test or one-way ANOVA).