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.

Gckr-P446L小鼠按PP、PL和LL基因型分为两组，一组在不含（-, 开放柱状图）或含（+, 填充柱状图）3 mg/kg AZD1656的高脂高糖饮食（HFHSD）下饲养20周。A）20周研究开始和结束时体重。B）AZD1656对Gckr mRNA和Gck mRNA（相对于18S的比例）无影响。C）8周龄P446L小鼠在常规饮食（RD）下的肝脏GKRP和GK免疫染色：LL小鼠中未见核GKRP染色和细胞质GK分布。D）P446L小鼠在常规饮食或20周HFHSD后的GKRP和GK代表性免疫印迹。E）在常规饮食或20周HFHSD研究后，LL小鼠中GKRP蛋白含量降低。F）在常规饮食或20周HFHSD研究后，LL小鼠中GK蛋白含量降低。G）在常规饮食或20周HFHSD研究后，LL小鼠中GK/GKRP免疫活性升高。H）20周HFHSD后的代表性GKRP和GK免疫染色。I-L）RD和20周研究结束时（n=4）GKRP和GK的核（N-H）和细胞质（C-H）H评分。I）LL小鼠中GKRP N-H评分降低。J）AZD1656降低LL小鼠中GK N-H评分。K）AZD1656增加PP（野生型）小鼠中GK C-H评分。L）AZD1656降低LL小鼠中GK核分布（N-H/C-H）。M）AZD1656增加PL和LL小鼠的肝脏GK活性。*P < 0.05为AZD1656的影响；#P < 0.05为基因型的影响。