vFLIP up-regulates MHC-I via activation of NF-κB. (A) MHC-I surface expression in LECs expressing vFLIP and the vector (pSIN). Mean and standard deviation were calculated after 3 independent infections (3 days after infection). (B) HLA-A mRNA levels of LECs expressing vFLIP and pSIN (3 days after infection). Expression is normalized to LECs infected with pSIN. (C) MHC-I expression in LECs treated with DMSO or BAY 11-7082 and then infected with vFLIP lentivirus or empty vector. MHC-I expression was determined 24 hours after infection and normalized to LECs treated with DMSO and infected with the empty lentiviral vector. (D) Surface expression of costimulatory class I proteins in vFLIP-expressing LECs. A 6-fold up-regulation of ICAM-1 is shown. (E) CD8⁺ T-cell proliferation after 7 days of coculture with LECs or LECs expressing vFLIP (see “Materials and methods” for details). Representative double staining (x-axis for CFSE and y-axis for CD8) dot plots are shown. The percent of cells in each quadrant is also shown. Average and standard deviation of CD8⁺ proliferated cells (CD8⁺/CFSE^low) from triplicate experiments is shown on the adjacent bar graph. Error bars were calculated based on standard deviation.