Figure 2D

A-C. Fibroblasts (MRC-5 or HDF) and ARPE-19 were transduced with lentivirus expressing UL88 or vector and subjected to puromycin selection. Cells were harvested for either WB or RNA analysis. A. MRC-5 cells expressing either vector or UL88 were analysed by western blot and probed for MyD88, UL88, and tubulin. Graph shows MyD88 protein levels quantified by densitometry analysis from 4 independent experiments. Results are shown as mean ± SD. *p<0.05. B. Cell lysates from HDFs expressing GFP vector, GFP-UL88, vector, or UL88, were analysed by western blot and probed for MyD88 and tubulin. C. Cell lysates from ARPE-19 cells expressing GFP vector or GFP-UL88 were analysed by western blot and probed for GFP, UL88, MyD88, and tubulin. D. Cells expressing either GFP-vector or GFP-UL88 were stimulated with IL-1β for 6 hours. Cells were fixed in PFA and stained for p65-NF-KB (red), GFP (green), and DAPI (blue). Cells were imaged on the confocal. E. At least ten different fields were images for each sample. The number of nuclei and NF-KB positive nuclei were counted. The percentage of NF-KB positive nuclei is graphed.