Fig. 8

The mTOR/GβL complex is a downstream target of the Ufl1/Ufbp1 complex. A IP analysis showing the interaction of Ufl1 or Ufbp1 with the mTOR/GβL complex. Tissue lysates derived from 2-month-old wild-type mice were immunoprecipitated using anti-Ufl1 or Ufbp1 antibody and immunoblotted with antibodies against GβL, mTOR, Ufl1 and Ufbp1. IP assays showing the interaction of GβL with the Ufl1/Ufbp1 complex. IgG served as a negative control in IP assays. B, C Western blot analysis showing that Ufbp1 overexpression deactivates mTOR signaling in human HCC Hep3B and HepG2 cells. D Histochemical analysis of liver tissues extracted from 7-month-old Ufbp1^Δ/Δhep mice treated with or without rapamycin. Black arrows indicate lipid-laden hepatocytes. E Immunohistochemical staining for GβL in 7-month-old Ufbp1^Δ/Δhep and control mice treated with or without rapamycin. Quantitative data are shown in the right panel (n = 5 mice/group). F Western blot analysis of the key molecules involved in mTOR signaling using the samples extracted from 7-month-old Ufbp1^Δ/Δhep and control mice treated with or without rapamycin (n = 5 mice/group). G Histochemical analysis of liver tissues extracted from 7-month-old Ufl1^Δ/Δhep mice treated with or without rapamycin. Black arrows indicate lipid-laden hepatocytes. H Western blot analysis of the key molecules involved in the mTOR signaling using the samples extracted from 7-month-old Ufl1.^Δ/Δhep and control mice treated with or without rapamycin. All western blots were independently repeated at least three times with consistent results. ** p < 0.01