Fig. 5. The temporary cytoplasmic p53 sequestration (7 dpi) was probably not caused by cytoplasmic tethering or the hyperactive nuclear export. HCMV-infected cells were treated with LMB, CHX or a combination of the two drugs. Incubation times are indicated in hours. All immunofluorescence images (A) were taken with the identical conditions, level of infections, IFA protocol, and equal exposure/digital picture processing conditions. Cells were stained as follows: anti-p53 (red, Texas Red-X), cytoplasmic anti-HCMV late proteins (green, FITC) and DAPI (blue). The corresponding western blot analysis (B) of the mock- and HCMV-infected cell extracts that were isolated from the identically infected cells treated with a single or a combination of the two drugs reconfirmed the IFA results shown in A. Numbers below the lanes are relative p53 protein levels and have been normalized against corresponding actin bands. (C) The pharmacological activity of LMB, 6 and 22 hours after use for the infected HUVECs treatment, was examined in fresh HUVECs. The same nuclear p53 accumulation pattern as those of the direct LMB treatment (data not shown) and increased p53 level confirmed that LMB remained pharmacologically active even 22 hours after incubation with the infected HUVECs.