Synergistic Activation of HIV-1 Expression by Deacetylase Inhibitors and Prostratin: Implications for Treatment of Latent Infection
Figure 6
Remodeling of nuc-1 after prostratin+VPA versus VPA treatment.
(A) Diagram indicating the positions of nucleosomes in the 5′ portion of the HIV-1 genome, the HinfI and AflII (*) cutting sites and the probe used in indirect end-labeling. An asterisk, representing the AflII cutting site, is located next to the band on the gel to permit its identification. (B) Nuclei were prepared from U1 cells mock-treated or treated with TNFα (10 ng/ml) (10 min), prostratin (5 µM), VPA (2.5 mM) and prostratin+VPA for 30 min, 1 h or 2 h and digested with AflII. After DNA purification and in vitro restriction with PstI, DNA samples were analyzed by indirect end-labeling using probe A (top panel) [93]. Size markers (a, b, c, d, e, f, g) have been previously described [93]. Quantification of the PstI-AflII bands was performed by radioimaging analysis using an InstantImager (Packard) (bottom panel) and results are presented as histograms indicating the band intensities relative to the intensity observed with mock-treated U1 cells, which was arbitrarily assigned a value of 1. Each value is the mean±SE of three separate experiments performed in duplicate.