Fig. 9
From: Expression of HIV from a 1-LTR circular DNA in the absence of integration
Number of copies of 1-LTR circles in transfected HeLa-tat cells in the presence or absence of aphidicolin. (A) HeLa-tat cells transfected with 1-LTRHIV and cultivated in the absence of aphidicolin. The number of copies of 1-LTR circles and the number of cells are plotted as a function of time in culture. The primer set used for the amplification of the 1-LTR circles consisted of: LA1 (forward), LA15 (reverse), and the probe 1-LTR-P. Detected DNA was normalized by measuring the amount of β-globin present in each sample with primers specific for the β-globin gene (see Materials and methods). The initial rapid decline in the number of 1-LTR amplicons is consistent with the disappearance of linear head to tail dimers co-formed by intermolecular ligation of 1-LTRHIV. (B) HeLa-tat cells transfected with 1-LTR DNA and incubated in the presence of aphidicolin. The number of copies of 1-LTR circles and the number of cells is plotted as a function of time in culture. (C) HeLa-tat cells transfected with 1-LTR and cultivated in the presence of aphidicolin. The increase in the number of 1-LTR circles and the amount of p24 detected in the culture supernatants are shown. (D) The amount of p24 released in the supernatant of HeLa-tat cells transfected with 1-LTRHIV in the presence or absence of aphidicolin is plotted as function of time in culture. Error bars represent one standard deviation from the average of two experiments, as described