Fig. 2

Functions of Sdf2l1 in vitro. a–c, f Cultured cells were infected with Ad-Ins^C96Y, whose lysates were immunoprecipitated for Western blotting: a, b Sdf2l1-floxed MEF cells, with a co-infection of Ad-Cre to knock out Sdf2l1, and with b further co-infection of Ad-Sdf2l1-FLAG to restore the expression; c NIH/3T3 cells, with either Sdf2l1 or Hspa5 knocked down, or with treatment with thapsigargin for 6 h; f primary hepatocytes, with Sdf2l1 or Tmed10 knocked down. In a, the lanes were run on the same gel but were noncontiguous. Multi: multiubiquitinated insulin, Mono: monoubiquitinated insulin. d RT-PCR to analyze ER stress marker gene expression in NIH/3T3 cells with Sdf2l1 and/or Hspa5 knocked down, treated with tunicamycin treatment (n = 3). e Wild-type mice were administered with Ad-Sdf2l1-FLAG intravenously (2.0 × 10⁷ PFU/g body weight [BW]), and refed for 6 h after fasting for 24 h, whose microsomal fractions were immunoprecipitated for Western blotting. g RT-PCR to analyze ER stress marker gene expression in primary hepatocytes, with ER-resident molecule(s) knocked down, treated with 100 nM insulin for 2 h after serum starvation for 16 h (n = 4). Values of the data are expressed as mean ± SEM. *P < 0.05, **P < 0.01. One-way ANOVA was used for assessment