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.

A) GK（葡萄糖激酶）在野生型446PP小鼠肝细胞中的转位作用（N/C，即细胞核/细胞质强度）由葡萄糖介导。数据为平均值 ± 标准误，涉及5个视野，1个肝细胞制备样本。
B-C) 在GKRP缺陷性肝细胞中，通过不同腺病毒滴度（5-40 x 10^6pfu/ml）表达的人hP、hL（B）或小鼠mP、mL（C）对葡萄糖磷酸化（[2-3H]葡萄糖代谢）的抑制作用。结果显示，在10-20 x 10^6pfu/ml时，hL或mL的抑制作用较低（# P < 0.05）。数据为平均值 ± 标准误，涉及3-12个肝细胞制备样本（B,C）。
D) 由GKOE相对于内源GK（实验细节参见图2 D，n=3个肝细胞制备样本）显著下调或上调（log-2变化）的基因列表，*表示这些基因已被芯片测序或基因微阵列识别为ChREBP靶基因。
E) 对由GK和GKRP相反调节的8个基因进行RT-qPCR验证（n=3个肝细胞制备样本）。