Figure 4

Figure 4

Dgat2 mutants produce fewer, abnormally smallApoB-containing lipoproteins.A, LipoGlo fish express ApoBb.1 with a C-terminal Nanoluciferase enzyme fusion (11)[Lipoprotein image re-used from (30) under the CC BY 4.0 license]. B, LipoGlo luminescence (RLU = relative luminescence units) in WT, dgat2^sa13945/+ and dgat2^sa13945 fish throughout embryonic development (2–6 dpf). Results represent pooled data from three independent experiments, n = 21 to 38 (WT), n = 65 to 75 (dgat2^sa13945/+), n = 32 to 43 (dgat2^sa13945) fish/genotype/time-point; mean ± SD. Significance was determined with a two-way ANOVA (overall p < 0.0001 for genotype) and Bonferroni’s multiple comparison test was performed to compare genotypes at each day of development (∗∗∗∗p < 0.0001 sa13945 versus WT and sa13945/+, ∗∗∗p < 0.001 sa13945 versus WT). C, representative LipoGlo PAGE gel of wild-type and dgat2^sa13945 mutants shows ApoB-containing lipoprotein (B-lp) size distribution from whole embryo lysates during development. B-lps are divided into four classes based on mobility, including zero mobility (ZM), very low density lipoproteins (VLDL), intermediate density lipoproteins (IDL) and low density lipoprotein (LDL). For heterozygote data, see original gels in Fig. S6. D, graphs show B-lp subclass abundance for WT, dgat2^sa13945/+ and dgat2^sa13945 fish at each day of embryonic development, analyzed from the gels in Fig. S6, as described in (11). Results represent pooled data from n = 9 fish/genotype/time-point; mean ± SD. For each particle class, significance was determined with a two-way ANOVA (overall p < 0.0280 ZM, p < 0.0001 VLDL, p < 0.0004 IDL, p < 0.0001 LDL for genotype) and Tukey’s multiple comparison test was performed to compare genotypes at each day of development (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001 sa13945 versus WT are shown).