FIG 6.

Effect of the DC474-480 mutations on UL37 interactions with gK. (A) Lysates of Vero cells infected with either wild-type HSV-1(F), VC1, or DC474-480 viruses were collected at 24 hpi, electrophoretically separated, blotted onto nitrocellulose membranes, and reacted with either anti-V5 (gK), anti-UL37, or anti-VP5 antibodies. The lanes in the blot for anti-UL37 and anti-V5 panels were spliced from different sections of the same blot, identified by the white line separating the lanes. (B) Lysates prepared as described for panel A were immunoprecipitated with the anti-V5 (gK) antibody. Electrophoretically separated proteins of the immunoprecipitates were blotted onto nitrocellulose membranes, and the presence of UL37 gK, UL20, or UL37 protein was detected by reacting the blots with either anti-V5, anti-FLAG, or anti-UL37 antibodies, respectively. No VP5 was detected when the immunoprecipitate was probed with anti-VP5 antibody (negative control).