Supporting Information(三)

来源：未知 2020-12-02 11:18

摘要：

　　Fig. S4. Menin is responsive to chemical carcinogen-induced liver chronic injury and inflammation. (A) The HE staining of the pancreatic islets in Men1WT and Men1+/- mice treated with DEN (25 mg/kg, i.p.) at 15 d of age and euthanized 8 mo

　　Fig. S4. Menin is responsive to chemical carcinogen-induced liver chronic injury and inflammation. (A) The H&E staining of the pancreatic islets in Men1WT and Men1+/- mice treated with DEN (25 mg/kg, i.p.) at 15 d of age and euthanized 8 mo after DEN injection. Original magnification was 100× or 200×, respectively.(B) The 6-wk-old Men1WT and Men1+/- male mice were injected with DEN (100 mg/kg, i.p.) and euthanized at 24 and 48 h after DEN exposure, respectively.Western blotting analysis was used for the activation of STAT3, AKT, and ERK1/2. (C and D) Circulating serum levels of serum IL-6 and TNFα were measured using ELISAs at the indicated time points. Error bars represent SEM (n = 3 or 4). (E) Male and female 6-wk-old Men1WT and Men1+/- mice were injected with CCl4 (2 mL/kg, i.p.) and euthanized at 24 and 48 h after CCl4 exposure, respectively. The expressions of menin, Yap1, pSTAT3, pAKT, AKT, and β-actin at the indicated time points were determined by Western blots. (F and G) Male and female WT C57BL/6 mice were injected with DEN (40 mg/kg, i.p.) at 3 wk of age.The pictures show representative damaged livers of male and female mice 4 mo after DEN injection. Liver lysates were used for immunoblot analyses with an antimenin antibody. (H) The tumor specimens (T) and corresponding surrounding tissue (N) from the DEN-induced livers were collected and lysed. Lysates were examined for expression and phosphorylation of the indicated proteins. (I) Menin and histone H3 ChIP assays were performed to evaluate the Yap1 promoters in DEN-induced HCCs (T) and adjacent tissues (N). Error bars represent SD (n = 3)

Fig. S5. The menin–Yap1 axis regulates IL-6 expression in liver. The WT C57BL/6 mice were treated with CCl4 (2 mL/kg, i.p.) at 6 wk of age and euthanized at 24 and 48 h after CCl4, respectively. (A) The mRNA levels of Men1, Yap1, IL-6, and TGF-β in the livers were assayed by qRT-PCR at 24 h. (B) Circulating serum levels of IL-6 were measured using ELISAs. (C) The ChIP assays of liver tissues were performed with an antimenin antibody at 48 h. (D and E) The HL-7702 and HepG2 cells were transfected with each of the three distinct siYap1 sequences, and the mRNA levels of Yap1 and IL-6 were determined using qRT-PCR. The relative mRNA levels were normalized to β-actin.

　　Fig. S6. Menin binds to the Yap1 promoter and influences the H3K4me3 modification in human HCC. Four cases of matched human HCC (T) and adjacent tissues (N) were analyzed by menin and H3K4me3 ChIP assays. The binding of menin and H3K4me3 modification at the Yap1 promoter were detected by qRTPCR. The menin in the four tumor specimens had been previously shown to be up-regulated by Western blots

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