Characterisation of C2cd4b-null mice. (a) C2cd4b global null mice (C2cd4bem2Wtsi) were generated by the IMPC. Using CRISPR/Cas9, the encoding exon from murine C2cd4b (exon 2) was deleted. (b) C2cd4b+/− (heterozygous) animals were setup as breeding pairs and the WT (C2cd4b+/+) and homozygous (C2cd4b−/−) littermates were studied. (c) RT-qPCR was performed on RNA from isolated islets and showed a significant decrease in C2cd4b mRNA levels in homozygous animals (p=0.0092). C2cd4b+/+n=4, C2cd4b+/−n=3, C2cd4b−/−n=7. **p<0.01, unpaired Student’s t test. (d, e) Changes in weight of C2cd4b+/+ and C2cd4b−/− female (d) and male (e) mice over time on an RC diet or HFD. RC: female (F)+/+n=3–6; F−/−n=4–13; male (M)+/+n=8–12; M−/−n=10–12. HFD: F+/+n=9–12, F−/−n=5–10–12; M+/+n=6–12, M−/−n=7–12. (f, g) Fasting blood glucose level in female (f) and male (g) mice on an HFD were measured at 23 weeks of age. F+/+n=9; F−/−n=5; M+/+n=5; M−/−n=7. (h, i) Percentage of body fat (h) and lean mass (i) in males maintained on an HFD at 20 weeks of age. *p<0.05, **p<0.01, ***p<0.001, mixed-effect analysis, RC-fed WT vs RC-fed mutant mice or HFD-fed WT vs HFD-fed mutant mice at each time point (d, e), or Student’s t test (c, f–i). Data were assessed for significance using an unpaired Student’s t test or two-way ANOVA where two genotypes were compared. Values represent means ± SEM
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