MolMet Figure 2: Functional assessment of human and mouse GKRP:P446>L by glucose-dependent translocation and transcriptome analysis.

A-C) Effects of glucose concentration (5-35mM) on nuclear sequestration (N/C: nuclear / cytoplasmic ratio) of GKRP (A) and GK (B),(C) in hepatocytes from GKRP-deficient mice transfected (10x10⁶pfu/ml) with human or mouse GKRP:446P/L (hP, hL, mP, mL) and cultured for 24h. A) Nuclear sequestration of GKRP is independent of glucose concentration. B) Translocation of GK at 35mM glucose in hepatocytes expressing human GKRP (hP or hL). C) Translocation of GK at ≥20mM glucose and ≥10mM glucose in hepatocytes expressing mouse GKRP mP or mL, respectively, n=3 hepatocyte preparations, *P<0.05 relative to 5mM glucose. D-H) Gene expression by unbiased RNA-transcriptome analysis in hepatocytes that were either untreated (none) or transfected (10x10⁶pfu/ml) with human (hP, hL) or mouse (mP, mL) GKRP and either without (GK-End) or with GK overexpression (GKOE) and cultured for 24h followed by 4h incubation with 25mM glucose before RNA extraction and processing for RNA-Sequencing, n=3 hepatocyte preparations (D). E) Gene counts for mouse Gck and Gckr showing expression by the adenoviral vectors. F) Numbers of significantly up-regulated or down-regulated genes by GKOE relative to the 5 combined groups of GK-End (none, hP, hL, mP, mL). G) Venn diagram showing genes down-regulated by GKRP with GKOE relative to GKOE alone. H) Gene counts for 14 of the genes up-regulated by GK and down-regulated by GKRP and for Hmgcr which is conversely regulated by GK and GKRP showing heterogeneity of response (hP, mP, hL, mL).