Fig. 6

Gm15441 suppresses Txnip protein expression through translational inhibition. A Western blot analysis of mouse primary hepatocytes transduced with adenovirus expressing YFP + empty vector, Txnip + empty vector, YFP + Gm15441-L, Txnip + Gm15441-L, or Txnip + Gm15441-S, for 24 h, MOI = 50. Quantitation is shown in the right panel; B Western blot analysis of Hepa1-6 hepatocytes transduced with adenovirus expressing YFP + empty vector, Txnip + empty vector, or Txnip + full length of Gm15441-L or one fragment of Gm15441-L (267–709 nt, 267–430 nt, 431–709 nt), for 24 h, MOI = 50. Quantitation is shown in the right panel; C representative fluoresces of RNA in situ hybridization staining of Gm15441 RNA (green) and Txnip RNA (red) in Hepa1-6 hepatocytes transduced with adenovirus expressing empty vector, or Txnip + Gm15441-L for 24 h, MOI = 50. All scale bars represent 5 μm; D quantitative real-time PCR analysis of pulled-down RNAs in Hepa1-6 hepatocytes transduced with adenovirus expressing Txnip and Gm15441-L for 24 h, MOI = 50, using non-specific oligo probe (control) and Gm15441 specific oligo probes; E Western blot analysis of Hepa1-6 hepatocytes transduced with adenovirus expressing Txnip + empty vector, or Txnip + Gm15441-L for 24 h, MOI = 50, followed by a 0, 20-min or, 40-min cycloheximide (CHX) treatment. Quantitation is shown in the bottom panel. Error bars are SEM, n = 2. *P < 0.05 (versus control), ^#P < 0.05 (versus Txnip + Adv group), N.S., not significant