FIG. 3.

Accumulation kinetics of total, integrated, and 2-LTR viral DNA forms following high-multiplicity infection of HuT-78 T cells. Infections were performed using 1 TCID₅₀ of HIV_HXB2 per cell with centrifugal enhancement. All PCRs were confirmed to amplify DNA in a linear fashion by quantification of standards (A to D) or dilution sets (E) (dilutions not shown). DNA markers (pUC19/HpaII) are indicated (M). (A) Total HIV DNA forms as measured by GAG-PCR using 500 cell equivalents of total DNA (combined Hirt supernatant and Hirt pellet). (B.i) Integrated HIV DNA levels as measured by LP-PCR on 100 cell equivalents of chromosomal DNA (Hirt pellet). (B.ii) Integrated HIV DNA levels as measured by the modified nested Alu PCR method performed on 1,000 cell equivalents of chromosomal DNA. (C.i) 2-LTR HIV DNA levels as measured by 2-LTR PCR on 500 cell equivalents of total DNA. (C.ii) Reanalysis of later time points for the 2-LTR DNA forms using 1,000 cell equivalents of total DNA. (D) β-Globin levels assayed by PCR on 50 cell equivalents of chromosomal DNA. Standards represent amplification of various amounts (based on cell counts) of HA8 chromosomal DNA. (E) Mitochondrial DNA levels assayed by PCR on 50 cell equivalents of Hirt supernatant (extrachromosomal) fraction.