MolMet Supp 2: Gckr transcript-1 is the predominant transcript expressed in mouse liver and mouse primary hepatocytes.
A-C) Relative abundance of Gckr transcripts-1,2, X1 and X2 in mouse tissues determined using primer sets targeting either a common sequence (spanning exons 14-15) or spanning the deletion sites of transcript-2, X1 and X2.
A) Relative selectivity of the 4 primer sets determined against RNA extracts from cells transfected with adenoviral vectors (10x106 pfu/ml) encoding the 4 transcripts: Gckr-t1 (RefSeq XM_006503881.3), Gckr-t2 (RefSeq BC012412), Gckr-X1 (RefSeq XM_006503882) and Gckr-X2 (XM_006503883).
B) Gck and Gckr mRNA are expressed at highest levels in liver.
C) Gckr-transcript-1 is the predominant transcript in liver and hepatocytes.
(B-C) Means ± SEM for n=4-8 extractions.
D) Amino acid alignment of mouse GKRP isoforms 1 and 2, predicted isoforms X1 and X2 versus human and rat sequences using Clustal Omega and visualized via Esprit3(Robert, X. and Gouet, P. (2014) "Deciphering key features in protein structures with the new ENDscript server". Nucl. Acids Res. 42(W1), W320-W324 - doi: 10.1093/nar/gku316). Showing deleted regions of Isoform 2, X1, and X2.
Funding
MICA: Exploring a new perspective on the mechanism of action of Glucokinase Activators in liver, a preclinical study
Medical Research Council
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