BP Figure 1: AZD1656 treatment increased liver GK activity in the P446L mouse.

Gckr-P446L mice of PP, PL and LL genotypes were fed on a high-fat high-sugar diet (HFHSD) without (-, open bar) or with (+, filled bar) 3 mg/kg AZD1656 for 20 wk. A) Body weight at the start and end of the 20 wk study. B) Lack of effect of AZD1656 on Gckr mRNA and Gck mRNA (ratio to 18S). C) Liver GKRP and GK Immunostaining in 8 wk old P446L mice on regular diet (RD): showing no nuclear GKRP staining and cytoplasmic GK distribution in LL mice. D) Representative immunoblots for GKRP and GK in P446L mice on either RD or after 20 wk on HFHSD. E) GKRP immunoactivity on RD or after 20 wk HFHSD study showing lower protein in LL mice. F) GK immunoactivity on RD or after 20 wk HFHSD study showing lower protein in LL mice. G) GK/GKRP immunoactivity on RD or after 20 wk HFHSD showing higher GK/GKRP in LL mice. H) Representative GKRP and GK immunostaining after 20 wk HFHSD. I-L) Nuclear (N-H) and cytoplasmic (C-H) H-scores for GKRP and GK on RD and at the end of the 20 wk study (n=4) I) Lower GKRP N-H scores in LL mice. J) AZD1656 decreased GK N-H scores in LL mice. K) AZD1656 increased GK C-H scores in PP (wild-type) mice. L) AZD1656 decreased GK nuclear distribution (N-H/C-H) in LL mice. M) AZD1656 increased liver GK activity in PL and LL mice. *P < 0.05 effect of AZD1656; #P < 0.05 effect of genotype.