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. Author manuscript; available in PMC: 2021 Jun 1.
Published in final edited form as: Hepatology. 2020 Feb 14;71(6):2118–2134. doi: 10.1002/hep.30966

Figure 7. SHP directly inhibits hepatic expression of miR-210 in part by repressing KLF4.

Figure 7.

(A) Hepa1c1c7 cells were transfected with siRNAs as indicated for 48 h. Levels of the indicated mRNAs (left) or miR-210 (right). (B) Hepa1c1c7 cells were transfected with luciferase vectors containing WT or mutated KLF4 motifs in the miR-210 promoter and with KLF4 and SHP expression vectors as indicated. Luciferase activity was normalized to that of β-galactosidase. (C) C57BL/6 (WT), SHP-KO, and FGF15-KO mice were refed for 6 h after fasting overnight and hepatic miR-210 levels were determined. (D-F) C57BL/6 mice were fed normal chow or CA-chow for 1 week. (D) Levels of hepatic Klf4 and Shp mRNA (left) or protein levels (right). (E) Nuclear localization studies: Levels of the indicated proteins in the nuclear and cytosolic fractions in liver extracts determined by IB. (F) Occupancy of indicated proteins at miR-210 promoter. Mean with SD (A-E) or SEM (F) is plotted. Statistical significance was determined by Student’s t-test (A, D, E), one-way ANOVA (B) or two-way ANOVA (C, F), n=5. * p < 0.05, ** p < 0.01. NS, not significant.