Figure 1.

Impact of HFM and microbes on the mouse intestine. (A) Experimental schematic highlighting microbial and nutritional conditions, genomic assays, and analysis. (B) Confocal en face images of jejunal villi 2 hours after gavage with egg yolk labeled with BODIPY C12 (red) with nuclei labeled with DAPI (blue). (C) Quantification of mean BODIPY C12 fluorescence per villus. Data points represent individual villi (27 GF+HFM villi, 42 CV+HFM villi), with open and closed circles representing villi from 2 biological replicate mice per condition. Averages and standard deviations of villi measurements are shown. Student t test showed significant differences when comparing across villi (P = .023) but not mice (P = .304). (D) Relative abundance of major lipid classes in each sample type including neutral lipids (triacylglyceride [TAG]), diacylglyceride [DAG], cholesteryl ester [CE]), phospholipids (phosphatidylcholine [PC], phosphatidylethanolamine [PE], phosphatidylinositol [PI], phosphatidylserine [PS]), and polar lipids (sphingomyelin [SM], ceramide [Cer], lysophosphatidylcholine [LPC], lysophosphatidylethanolamine [LPE], hexosylceramides [HexCer]). All measurements were normalized to internal standards and are shown as fold change relative to GF. Two-way ANOVA revealed there was not a statistically significant interaction between treatment and lipid class (F_33,144 = 0.8728, P = .6672). Simple main effects analysis showed that treatment did have a statistically significant effect on lipid class abundance (P = .0367). (E) Percentage of FAs detected across all neutral lipid classes (TAG, DAG, and CE) that are saturated FAs (SFA), monounsaturated FAs (MUFA), and polyunsaturated FAs (PUFA). Two-way ANOVA revealed there was a statistically significant interaction between treatment and FA saturation group (F_6,36 = 13.93, P < .0001). Simple main effects analysis showed that treatment did not have a statistically significant effect on lipid class abundance (P > .9999). (F) Percentage of FAs detected across all neutral lipid classes (TAG, DAG, and CE) with the corresponding chain length and saturation. Two-way ANOVA revealed there was a statistically significant interaction between treatment and FA type (F_75,312 = 10.98, P < .0001). Simple main effects analysis showed that treatment did not have a statistically significant effect on FA abundance (P > .9999). For the data shown in (D-F), significant differences (P < .05) by post hoc Tukey multiple comparisons tests are noted for (a) GF vs CV, (b) GF+HFM vs CV+HFM, (c) GF vs GF+HFM, and (d) CV vs CV+HFM. Data are shown as average and standard deviation of 4 mice per condition. See also Supplementary Table 1.