Fig. 10. Cytoplasmic loading onto MHCII is insensitive to reduction of PI 3-kinase activity and 3-methyladenine treatment. BMC-2 cells were transfected and treated with autophagy inhibitors as indicated to the right of each plot (3MA, 10 mM; LY294002, 50 µM). The generation of Y-Ae⁺ peptide MHCII complexes was assayed by flow cytometry at 20 hours post-transfection. The numbers in the top right quadrant indicate that the percentage of eGFP-expressing cells that are also positive for Y-Ae⁺ complexes is unaffected by the different treatments tested. Numbers in the top left quadrant indicate percentage of eGFP-expressing cells; numbers in the bottom right quadrant indicate percentage of cells gated for positive Y-Ae signals; numbers in the bottom left quadrant indicate percentage of unstained cells.