Figure 3

Modulating lipid metabolism influences LD levels in macrophages. RAW264.7 cells were treated with 10 µM C75 for 24 h to inhibit fatty acid synthesis, incubated with 360 µM oleic acid for 24 h, and infected with S. uberis in mid-exponential phase (MOI = 10) for 3 h at 37 ℃. A BODIPY 493/503 staining for LD visualization (green) in different groups. Images are representative of n = 10 samples/group. Scale bar, 20 µm. B, C Flow cytometry of BODIPY-stained RAW264.7 cells. Left: Representative histograms. Right: Mean fluorescence intensities from 3 independent experiments. The dotted line indicates background fluorescence in uninfected cells. Data are presented as the means ± SEMs (n = 6). *(P < 0.05) = significantly different between the indicated groups by one-way ANOVA. D–F NEFA, TG and TC contents in RAW264.7 cells during S. uberis challenge. Experiments B–F were conducted in triplicate. Data are presented as the means ± SEMs (n = 6). *(P < 0.05) = significantly different between the indicated groups by one-way ANOVA.