FIG. 6.
Effects of HSV infection on the phosphorylation status of Src family kinases in HEL cells 6 h after infection. Replicate cultures of HEL cells grown in T150 flasks were mock infected or exposed to 10 PFU of HSV-1(F), ΔICP0, ΔUS3, ΔUL13, or ΔUS3/ΔUL13 viruses per cell. The cells were harvested at 6 h and lysed as described in Materials and Methods. For each sample, 1 mg clarified whole-cell lysate was reacted with 3 μg of anti-Src antibody for 3 h at 4°C. The immune precipitates were resuspended in 120 μl of SDS disruption buffer, one-third of which was separated in a denaturing polyacrylamide gel, along with 70 μg of total cell lysate from each sample. The electrophoretically separated proteins were reacted with the anti-Src-pY416 or anti-Src-pY527 antibodies.