Supplementary Figure 4: Clinical chemistry analysis of heterozygous Klf14tm1(KOMP)Vlcg knockout mice.
Clinical chemistry parameters were measured in male knockout C57BL/6N Klf14 mice and their wildtype controls. Since Klf14 is mono-allelically maternally expressed in mouse and human, we compared heterozygous mice that had inherited the deletion allele from their mother (heterozygous-MAT, expressing the deletion) with heterozygous mice that had inherited the deletion allele from their father (heterozygous-PAT, not expressing the deletion). We also compared the two groups to their own homozygous wildtype colonymate controls (wildtype-MAT and wildtype-PAT). This was because we used two separate crosses to produce the MAT and PAT carrier cohorts; one stock from heterozygous mothers and one from heterozygous fathers, each crossed to wildytpe C57BL/6N mice. Mice were fed a standard diet and then switched at 18-weeks of age to a 45kcal% high fat diet. Comparing across the timecourses for (a), HDL-C; (b), insulin; (c), glucose; (d), IPGTT by calculating area under the curve, base-lined to t=0 values (data not shown), and analysing with a 1-way ANOVA non-parametric Kruskal-Wallis test and Dunns multiple comparison test we found: that HDL-C (a) was lower in MAT compared to either PAT or wildtype-MAT (p= 0.0055 and 0.0086 respectively) and that wildtype-PAT compared to PAT was not significantly (p=0.56) different; that insulin (b) was lower in MAT compared to either PAT or wildtype-MAT (p= <0.0003 and 0.0020 respectively) and that wildtype-PAT compared to PAT was not significantly (p>0.99) different; that Glucose (c) was lower in MAT compared to PAT (p= <0.0001) and that wildtype–MAT and wildtype-PAT compared to MAT and PAT respectively were not significantly (p=0.79 and >0.99 respectively) different; and that for an IPGTT (d) none of the group comparisons were significantly different (p>0.99) to each other. Then to examine effects at specific times for (a) HDL-C, (b) insulin, (c) glucose and (d) IPGTT individual pairwise comparisons were made at each timepoint using a Mann-Whitney 2-tailed t-test and were significantly reduced in the MAT group compared to PAT group at 8, 22 and 27 weeks in (a,b,c) and at all timepoints in (d). The MAT groups were also significantly lower compared to wildtype-MAT for HDL-C and insulin at 22 and 27 weeks. Values are expressed as mean ± SD and in (a,b,c) wildtype-MAT n=16, wildtype-PAT n=9, heterozygous-MAT n=15, heterozygous-PAT n=16 and in (d) wildtype-MAT n =19, wildtype-PAT n=10, MAT n=16, PAT n=16. For (e), HDL-C; (f), LDL-C and (g), total cholesterol was measured in a 33-week blood sample collected under terminal anaesthetic and was significantly, using an unpaired 2-tailed t-test, reduced in heterozygous-MAT mice compared to PAT mice. Values are expressed as mean ± SD (PAT N =8, MAT N =8). Wildtype MAT grey, wildtype PAT black, MAT KO red, PAT KO blue lines and fill
