Fig 7.

A26 protein in vitro binds to A16 and G9 proteins that are not in complex with A56/K2. (A) GST-A26 pulled down A16 and G9 proteins but not A56 protein. HeLa cells were infected with WR, WRΔA56R, and WRΔK2L at an MOI of 5 PFU/cell; cell lysates were harvested at 24 h p.i. for GST pulldown analyses with 30 μg of recombinant GST and GST-A26 proteins and analyzed by immunoblotting with anti-A16 (1:1,000), anti-G9 (1:5,000), and anti-A56 MAb (B2D10) (1:500). (B) GST-A26 did not pull down K2 protein. HEK293T cells were transiently transfected with plasmids expressing HA-A16, Myc-G9, A56-GFP, and K2-Flag proteins individually or in combination. The lysates were harvested for GST pulldown and analyzed by immunoblotting as described for panel A with anti-HA (1:1,000), anti-Myc (1:1,000), anti-GFP (1:4,000), and anti-Flag (1:1,000) antibodies. Input represents 1% of total cell lysates.