Figure 3.

Loss of DGAT1 results in decreased lipid droplet formation in organoids. (A) Immunofluorescent images of 4′,6-diamidino-2-phenylindole (DAPI) (blue) and LD540 (yellow) staining of organoids from healthy control, DGAT1 mutant patient 8 (P8) and DGAT1^KO organoids after 17-hour incubation with vehicle control (BSA), 1 μM OA, or 1 μM OA + 0.1 μM DGAT1 inhibitor (OA+DGAT1i). Representative images of 3 healthy controls, 3 patients (patients 7–9), and 3 CRISPR/Cas9 genome-edited DGAT1-knock-out (DGAT1^KO) organoids. (B) Representative histograms of SSC and LD540 staining in organoids from controls, patients, and DGAT1^KO organoids as described in (A). Upon OA stimulation, control organoids accumulate lipid droplets and show increased SSC and LD540, which was severely reduced in patient-derived and DGAT1^KO cells. Mean fluorescence intensity (MFI) of SSC and LD540 was plotted for n = 3 per group. Statistical analysis was done using a 2-way analysis of variance with Tukey’s multiple comparison test. Mean ± SD is indicated; * P ≤.05, *** P ≤.001.