Critical Role of HAX-1 in Promoting Avian Influenza Virus Replication in Lung Epithelial Cells

Figure 4

The N-terminus of PB1-F2 was critical for its interaction with HAX-1. (a) Interaction of PB1-F2 and HAX-1 in yeast cells. Yeast cells were cotransformed with pGBKT7-PB1-F2 expressing a full length of 1–90 residues and a segment of the N-terminal 1–60 residues or C-terminal 30–90 residues, together with pGADT7-HAX-1 (full length). One week later, growing clones on each SD/dropout plates were propagated and validated on X-α-Gal/SD/dropout plates. Clones generating blue indicate positive interaction, whereas white color indicates negative interaction. (b) Colocalization of PB1-F2 and HAX-1 in the mitochondria of A549 cells. A549 cells were transfected with empty vectors (not shown) or transfected with plasmids expressing HAX-1 or/and PB1-F2 for 24 hrs, subjected to immunofluorescence staining with respective antibodies. The cells were costained with MitoTracker (middle and bottom panels).

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