Expression of nuclear transcription factors during the PTT of HFD-fed WT and AnxA6-KO mice.

(A) Nuclear fractions from liver samples from WT and AnxA6-KO (AnxA6-/-) mice before (0 min; WT lane 1–4, AnxA6-KO lane 5–8) and 120 min after pyruvate administration (120 min; WT lane 9–12, AnxA6-KO lane 13–16; n = 4 per group) were prepared and analyzed by western blotting for the transcription factors FoxO1, PPARα, SREBP1 and LXR. β-actin served as loading control. Molecular weight markers are shown. Arrowhead points at LXR. (B) Relative levels of FoxO1, PPARα, SREBP1 and LXR were quantified and normalized to β-actin expression. The mean values (± SEM) relative to WT at t = 0 min are shown. (C) RNA from HFD-fed WT and AnxA6-KO livers before (0 min) and 120 min after pyruvate administration was isolated (n = 4 per group). cDNA was generated and RT-PCR for AnxA6, fibroblast growth factor 21 (Fgf21), glucose-6 phosphatase (G6P) and insulin induced gene 1 (Insig1) was performed as described in Material and Methods. Relative mRNA expression was normalised to the housekeeper Tbp levels using the ΔΔCT method. The expression relative to the WT at t = 0 min is shown. * P < 0.05, ** P < 0.01, *** P < 0.001 (Student’s T-Test).