Fig 2: PODXL decreases the expression of CD86, MHC-I and MHC-II in APCs. (A) Raji-PODXL, Raji-Ctrl cells and CMTMR-labeled Jurkat cells were preincubated separately with a blocking anti-LFA-1 monoclonal antibody for 15 min. Afterwards, Raji-PODXL and Raji-Ctrl were mixed with CMTMR-labeled Jurkat cells at a ratio of 1:1 and incubated for 30 min. The percentage of Raji-Jurkat cell conjugates was determined by flow cytometry as in ( Figure 4A ). Bar graph represents the mean ± SD of nine independent experiments. *p < 0.05, ****p < 0.0001, one-way ANOVA. (B) Raji cells overexpressing PODXL and Raji Ctrl cells were analyzed by flow cytometry for surface expression of CD54, CD80, CD86, MHC-I and MHC-II. Overlay histograms represent the surface expression of the indicated marker in Raji-PODXL cells (red line), Raji-Ctrl cells (blue line) and the isotype control (grey line). Graphs show mean ± SD of surface expression (?MFI= Marker MFI - isotype control MFI) from six (CD54, CD80, CD86, MHC-I) or five (MHC-II) independent experiments. *p < 0.05, **p < 0.01, paired t test. 0.0001, ns, not statistically significant; one-way ANOVA.