Whole-mount imaging of apolipoprotein B-containing lipoprotein (ApoB-LP) localization using LipoGlo chemiluminescent microscopy. a Representative images of ApoB-LP localization patterns from analysis of 15 larvae per genotype from wild-type (WT), mtp−/−, and apoC2−/− genotypes (6 days post fertilization (dpf)). The white arrow and arrowhead mark the larval intestine and liver respectively. b LipoGlo signal colocalizes with the central nervous system marker Tg(Xla. Tubb2-mApple-CAAX), quantification in Supplementary Fig. 5. c LipoGlo signal localized to subregions of the central nervous system (CNS). d–f Representative images and g–i quantification of ApoB-LP localization across developmental, genetic, pharmacological, and dietary manipulations. g Signal localization at each day of larval development in WT (degree of freedom (DF) = 5, n = 15, Welch’s analysis of variance (ANOVA) p < 0.0001 for each region over time), mtp−/− (DF = 11, n = 15, two-way robust ANOVA p < 0.001 for all regions, Games–Howell p < 0.001), and apoC2−/− (DF = 11, n = 15, two-way robust ANOVA was not significant for any region) genetic backgrounds. Upward-facing arrowheads (green) indicate significant enrichment of that species at that time point compared to WT, and downward-facing arrowheads (magenta) indicate depletion using the Games–Howell test with a threshold of p < 0.05. h Signal localization from 3 to 5 dpf in larvae treated with 10 μM lomitapide or vehicle control (DF = 11, n = 15, two-way robust ANOVA p < 0.001 for head and viscera, Games–Howell p < 0.0001). i Subclass abundance from 10 to 15 dpf in larvae subjected to a fasting and re-feeding paradigm. The first bracket delineates changes relative to time 0 (the onset of the fasting period), and the second bracket delineates changes relative to time point 48 (the onset of the re-feeding period) (DF = 10, n = 15, Welch’s ANOVA p < 0.0001 for each region, Games–Howell p < 0.005). Supplementary Figure 4 displays standard deviations for g–i. Results represent pooled data from three independent experiments, “n” denotes number of samples per data point. Scale bars = 500 μm.
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