Fig. 6.
Autophagy is not a significant pathway of MHC class II-mediated presentation during influenza viral infection in dendritic cells. BMDCs from BALB/c or CB6F1 mice were transduced with LMP alone or LMP-ATG7 shRNA retrovirus. (A) Forty-eight to 72 h after transduction cells were harvested and sorted for high GFP expression (GFPhi), and knockdown was assessed by Western blotting. (B) Functional knockdown of autophagy was determined by probing infected cell lysates for p62 accumulation via Western blotting (top, raw data; bottom, densitometry analysis). (C) GFPhi BMDCs were infected with PR8 and cultured with CD4+ T cells purified from influenza virus-primed mice overnight in an IFN-γ ELISpot assay (left, 48 h knockdown; right, 72 h knockdown). Results are representative of at least two independent experiments at each time point.