Fig. 5.

Toledo wild-type (WT) and ToledoΔUL133-138 express IE, E, and L genes with similar kinetics. Primary fibroblasts were infected with Toledo WT or ToledoΔUL133-138 as indicated. (A) Cells were harvested at the indicated time points, fixed in ethanol, and stained with IE1/IE2 antibodies to detect IE gene expression, an anti-gB antibody to assess expression of a representative of early gene expression, and an antibody directed against pp28 to analyze late gene expression. Alexa Fluor 488 fluorescence was measured by flow cytometry. Cells were harvested and lysed, and protein extracts were prepared. Viral protein expression was detected by immunoblotting using IE-, gB-, and pp28-specific antibodies. Equal protein loading was controlled for by including GAPDH.