FIG. 2.
VP22 is important for inhibiting CD1d recycling upon an HSV-1 infection. HEK293-CD1d cells were mock treated (gray-filled histogram) or infected with WT HSV-1 (solid line), ΔUL49 (dotted line) or UL49R (dashed line) viruses at an MOI of 2 for 4 h (A). The cells were fixed, permeabilized with 0.1% saponin, stained with an anti-HSV antibody, and analyzed by flow cytometry. (B) The same cells that were mock treated (open circles), or infected with WT HSV-1 (open triangles), ΔUL49 (solid circles), and UL49R (solid triangles), were blocked with purified anti-CD1d (42.1) and then cultured at 37°C as described in the Materials and Methods section. At each indicated time point, a cell aliquot was harvested and stained with PE-labeled 42.1 mAb. The level of recycled CD1d is shown and was calculated by subtracting the mean fluorescence intensity (MFI) at each time point from time 0. ns, not significant; data shown are representative of three independent experiments. ****p < 0.0001 (two-way ANOVA). PE, phycoerythrin.