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. Author manuscript; available in PMC: 2014 Dec 20.
Published in final edited form as: Nat Commun. 2014 Jun 20;5:4233. doi: 10.1038/ncomms5233

Figure 1. Obesity induces Sestrin2 through ER stress signaling.

Figure 1

(a-c) HepG2 cells were treated with palmitic acid (PA) for indicated hr and analyzed by immunoblotting (images with black bands) and RT-PCR (images with white bands) (n = 3). P values were calculated between untreated (0 hr) and indicated groups (b,c). (d,e) At 48 hr after infection with GFP (Con)- or SERCA2b-overexpressing adenoviruses, cells were treated with BSA (0 hr) or PA for indicated hr (d) or 9 hr (e) (n = 3). (f,g,l,m) 6-month-old WT mice kept on LFD or HFD for 4 months were injected daily with vehicle (PBS) or TUDCA (500 mg per kg body weight i.p.) or transduced once with adenoviruses expressing SERCA2b, GFP (Con) or shRNA targeting c/EBPβ. After 10 days, livers were analyzed by immunoblotting (n = 3). (h,k) At 48 hr before PA treatment, cells were infected with lentiviruses expressing shRNAs targeting luciferase (Con), PERK (h) and c/EBPβ (k). After 9 hr of BSA (−) or PA treatment, cells were analyzed by immunoblotting. (i,j) Chromatin immunoprecipitation (ChIP) analysis of cells with indicated treatments (n = 3). (n) At 24 hr after infection with GFP (Con)- or c/EBPβ-overexpressing adenoviruses, cells were analyzed by immunoblotting. All data are shown as the mean ± s.e.m. P values are from Student’s t test. Molecular weight markers are indicated in kDa (immunoblots) or bp (agarose gels).