MolMet Figure 3: Lower GKRP and GK protein levels in human NAFLD biopsies of the rs1260326-TT genotype and in the P446L mouse.

A-I) Human NAFLD biopsies genotyped for GCKR rs1260326 variant (CC n=23; TT n=17) and scored as simple steatosis (NAFL) or NASH were stained for GKRP or GK and nuclear (N-H) and cytoplasmic (C-H) H-scores were determined. (A) representative images. (B-D) Lower GKRP N-H scores by TT genotype for entire cohort (B) or by -/+ NASH diagnosis (C), lower C-H scores (D) (E-G) Lower GK N-H scores by TT genotype for entire cohort (E) or by -/+ NASH diagnosis (F) and lower C-H scores (G) H) Correlation of GK N-H vs GKRP N-H, r2 0.34, P&lt;0.0001 for entire data set. I) Lower GK (N-H) / GKRP (N-H) ratio by TT genotype, # P&lt; 0.0003. J-M) GKRP P446L mouse aged 14-wk (regular diet, n=4,4, PP, LL) or 28-wk (on HFHSD for last 20-wk, n=10,10, PP, LL): Liver <em>Gckr</em> and <em>Gck</em> mRNA normalized to RplpO (J); GKRP and GK protein (K-L) and GK enzyme activity (M). J) Similar <em>Gckr</em> mRNA by genotype but lower <em>Gck</em> mRNA in 28-wk mice by LL genotype. K-L) Lower GKRP and GK immunoactive protein (n=4-8) and lower GK activity (M, n=4,4,10,10,10) by LL genotype. #P&lt;0.05, LL vs PP; *P&lt;0.05 28-wk vs 14 wk. N-R) Comparative GKRP and GK immunostaining of human liver biopsies (CC,TT: n=7,6) and P446L mouse liver (PP, PL, LL: n=7,4,5) showing representative images for mouse (N) and the N-H and C-H scores. O) Lower GKRP N-H scores for human compared with mouse ($) liver and by TT and LL genotypes (#) and negligible cytoplasmic H-scores. P) Lower GK N-H and C-H for human compared with mouse liver ($) and lower N-H by TT (human) or LL (mouse) genotype (#). Q) Correlation of GK and GKRP N-H scores. R) Relative distribution of GK in nucleus and cytoplasm (N-H/C-H ratio) showing lower nuclear GK sequestration by TT and LL genotypes. #P&lt;0.05 by genotype; $P&lt;0.05 human vs. mouse.