Fig. 7: In vivo role of GABP in mice.
From: Non-human primate seasonal transcriptome atlas reveals seasonal changes in physiology and diseases
a CRISPR/Cas9 targeting of the mouse Gabpb1 exon 9. Schematic representation of the mouse Gabpb1 gene illustrates crRNAs (blue characters) and the protospacer adjacent motif (PAM) sequences (red characters) tailored to exon 9. Sequencing of the deletion band shows ligation of the Cas9 target site. PCR across the genomic deletion region shows the smaller PCR product in mutants. Western blotting of Gabpβ1 with an antibody recognizing both long and short forms shows absence of long form of Gabpβ1 in the mutant homozygous (Mut/Mut) mice. β-actin was detected as loading control. b Effects of 1-week exposure to SC or LW conditions on Gabpb1 expression and organ weight in Gabpb1 exon 9 deletion mutant heterozygous mice and their wild-type littermates (two-way ANOVA, Šídák’s multiple comparisons test, mean + SEM, +/+: n = 3; +/Mut: n = 4). Numbers within the graph indicate the P value. c Immunohistochemistry of Ki67 in the heart (HEA), pancreas (PAN), and kidney (KID). In WT mice, the number of Ki67 immuno-positive cells (red arrows) was higher under SC than LW conditions in the HEA and PAN, while glomerular size was larger under SC than LW conditions in the KID. These differences were smaller in Gabpb1 heterozygous mutant mice. L: Islet of Langerhans; G: Glomerulus. d H&E staining of cardiomyocytes. Changes in cardiomyocyte diameter between SC and LW conditions were more obvious for WT than Gabpb1 heterozygous mice. e Immunohistochemistry of β-catenin, which localizes to the acinar cell membrane. Changes in the size of pancreatic acinar cells between SC and LW conditions were more obvious in WT than Gabpb1 heterozygous mice. Scale bars: 50 μm. f Effects of 1-week exposure to SC or LW conditions on Ki67-positive cell numbers, cardiomyocyte diameter, pancreatic acinar cell size, and glomerular size in Gabpb1 exon 9 deletion mutant heterozygous mice and their wild-type littermates (two-way ANOVA, Šídák’s multiple comparisons test, mean + SEM, +/+: n = 3; +/Mut: n = 4). Numbers within the graph indicate the P value.
