FIG 2.

Analysis of pCREB expression in VZV-infected fibroblasts and effects of inhibition of pCREB transcriptional activity. (A) HELFs were infected with VZV and treated with either 0.1% DMSO or 1 μM XX-650-23 at the time of infection. Twenty-four hours postinfection, cytoplasmic and nuclear fractions were prepared. The lysates were analyzed by Western blotting with antibodies against VZV IE62 (control for infection), pCREB, alpha-tubulin (cytoplasmic protein control), and lamin A/C (nuclear protein control). Whole-cell lysates from forskolin-treated SK-H-MC human neural cells (Sigma) were included as a control for the detection of pCREB. Quantification was done using ImageJ software; the intensities of the bands detected with anti-pCREB antibody were collected and normalized to the intensities of the bands detected with anti-alpha-tubulin for cytoplasmic extracts and with anti-lamin A/C for nuclear extracts. (B) Fibroblasts were left uninfected or inoculated with VZV or UV-inactivated VZV. Extracts were prepared 24 h postinfection and analyzed by Western blotting to detect IE62, pCREB, and alpha-tubulin as a control. (C) Lysates prepared at 24 h were analyzed by Western blotting for cyclin D1 and cyclin D2. Alpha-tubulin was used as a control for cell proteins and IE62 as a control for infection. UI, uninfected; I, infected; ctrl, control. Numbers to the left of each blot are molecular masses (in kilodaltons).