(A) Electron micrographs of mouse primary adipocytes 24 hr post-mock or Toxoplasma infection. Toxoplasma vacuoles are pseudocolored red. Scale bar, 5 μm.

(B) Dot plot of the average area of an individual lipid droplet (LD) in n = 40 cells in experiments as in (A). Means ± SEM are plotted. ∗∗∗∗p < 0.0001 in an unpaired t test.

(C) Confocal images of mock- and Toxoplasma RFP+ (red)-infected Caco-2 cells labeled with BODIPY493/503 (green) and imaged 24 hr post-infection (hpi). Selected cells are contoured by a dashed line in merged panels. Scale bar, 25 μm.

(D) Dot plot of LD number in experiments as in (C) in n = 200 cells. Means ± SEM are plotted. ∗∗∗∗p < 0.0001 in an unpaired t test.

(E) Schematic representation of fluorescent FA-pulse and infection-chase assay. MEFs were incubated overnight with FL C 12 , rinsed, mock- or Toxoplasma-infected, and analyzed by flow cytometry or fluorescence microscopy.

(F) Combined bright-field epifluorescence images of FL C 12 -pulsed cells at 24 hpi. Representative regions of interest (ROIs) used to quantify the mean fluorescence intensity (MFI) are outlined in black (uninfected cells cytoplasm), blue (infected cells nuclei), or gray (Toxoplasma vacuoles). Scale bar, 10 μm.

(G) Mean ± SEM from n = 3 independent experiments of relative FL C 12 accumulation in ROIs of uninfected cells cytosol or Toxoplasma vacuoles in experiments as in (F). ∗∗p < 0.01, ∗∗∗∗p < 0.0001 Toxoplasma vacuole versus uninfected cytosol, #p < 0.001 6 hpi versus 24 hpi by two-way ANOVA.