Composite of autoradiograph, immunoblot, and immunofluorescence images. HCMV encodes a genetic mechanism expressed in the initial hours of infection to block phosphorylation of eIF-2α and maintain protein synthesis. (A) An autoradiographic image of the electrophoretically separated pulse-labeled proteins from mock-infected (Mock), HCMV-infected (HCMV), or UV-inactivated HCMV-infected (UV) HFF cells subjected to further mock or R3616 superinfection. (B) Immunoblot image shows abundant IE1 immunostaining in the cells infected with HCMV alone and reduced staining in the HCMV/R3616-infected cells. (C) Immunoblot showing equivalent ICP0 immunostaining in the R3616-infected cells. (D) Immunoblot image demonstrates abundant phosphorylated eIF-2α in both mock/R3616 and UV-inactivated HCMV/R3616-superinfected cells but minimal levels in the HCMV/R3616-superinfected cells. (E) Immunoblot demonstrating equivalent levels of total eIF-2α in the infected cell samples. (F) Indirect immunofluorescence of mock-infected (Mock), HCMV-infected (HCMV), and UV-inactivated HCMV-infected (UV) cells. The first row shows detection of virion-associated protein pp65 (pUL83) in both HCMV- and UV-HCMV-infected cells. The second row of images demonstrates the presence of IE1 staining only in HCMV-infected cells. The third row shows Hoechst nuclear staining in all samples.