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

<p>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.</p> <p>A) Nuclear sequestration of GKRP is independent of glucose concentration.</p> <p>B) Translocation of GK at 35mM glucose in hepatocytes expressing human GKRP (hP or hL).</p> <p>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.</p> <p>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).</p> <p>E) Gene counts for mouse Gck and Gckr showing expression by the adenoviral vectors.</p> <p>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).</p> <p>G) Venn diagram showing genes down-regulated by GKRP with GKOE relative to GKOE alone.</p> <p>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).</p>