MolMet Supp 5: Complementary data to Figure-2: GK translocation; GKRP-inhibition of glucose phosphorylation; Differentially expressed genes by GK overexpression from RNA-sequencing and RT-qPCR validation.
A) Translocation of GK by glucose (N/C, nuclear/cytoplasmic intensity) in wild-type 446PP mouse hepatocytes. Mean ± SEM, 5 fields, 1 hepatocyte preparation.
B-C) Inhibition of glucose phosphorylation (metabolism of [2-3H]glucose) by human hP, hL (B) or mouse mP, mL (C) GKRP expressed in GKRP-deficient hepatocytes over a range of adenoviral titres 5-40 x 106pfu/ml showing lower inhibition by hL or mL (# P < 0.05) at 10-20 x 106pfu/ml. Means ± SEM for 3-12 hepatocyte preparations (B,C).
D) List of genes significantly down-regulated or up-regulated (log-2 change) by GKOE relative to endogenous GK (experimental details as in Figure 2 D, n=3 hepatocyte preparations), *denotes genes previously identified as ChREBP target genes by CHIP-sequencing or gene microarrays.
E) RT-qPCR validation (n=3 hepatocyte preparations) of 8 genes conversely regulated by GK and GKRP.
MICA: Exploring a new perspective on the mechanism of action of Glucokinase Activators in liver, a preclinical study
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