FIG 3.

MDV infection increases the formation of neutral lipid droplets. (A) Visualization of cytoplasmic lipids in neutral lipid droplet organelles of CEF cells in MDV-infected (i) and mock-infected (ii) cells (magnification, ×10) at 72 hpi. Black arrows indicate lipid droplets. (B) Confocal microscopy imaging with maximum projection of z-stacks for each channel demonstrating nuclear and cytoplasmic distribution of pRB1B UL35-GFP virus (green) and lipid droplets (red). Mock- and pRB1B UL35-GFP-infected cells were fixed at 72 hpi and stained with DAPI (nuclear stain) and the neutral lipid stain LipidTOX-568nm. Images 5 and 10 are three-dimensional representative images analyzed using IMARIS software. z-stacks were analyzed using the IMARIS spot function analysis tool to quantify the relative amount of lipid droplets per cell in MDV-infected and mock-infected CEFs (50 to 90 cells). (C) The numbers of lipid droplets per cell in MDV-infected and mock-infected CEFs at 72 hpi. (D) The numbers of lipid droplets per cell in MDV-infected CEFs treated with TOFA (1.54 μM) or C75 (5.9 μM) are shown. ****, statistically significant difference compared to the control (P < 0.0001). All experiments were performed in duplicates, and data are representative of 3 independent experiments.