HIV-TCM increases VEGF-A expression in HUT 78 cells. (A) Northern blot analysis of VEGF-A mRNA expression in HUT 78 cells. Cells were incubated in normal cell culture medium (control) with TCM or HIV-TCM. In two separate experiments, total RNA was isolated after 1, 3, and 5 days of incubation (left) or after 5 days of incubation (right), subjected to electrophoresis (20 μg per lane), and blotted onto a nylon membrane. Hybridization was performed with a radioactively labeled human VEGF-A specific cDNA probe (specific activity, ≥8 × 108 cpm/μg DNA) at 42°C for 16 hours. VEGF-A mRNA expression in HUT 78 cells increased by twofold after 3 and 5 days of incubation with HIV-TCM as compared with TCM and threefold as compared with control cells incubated in normal medium. (B) Ethidium bromide staining of blotted RNA indicated that equal amounts of RNA were used in each lane. (C) Densitometric evaluation of band intensities from the Northern blot shown in (A). RNA derived from cells incubated with TCM or with HIV-TCM were blotted and hybridized on the same filter. VEGF-A expression was similar after 1 day of incubation with HIV-TCM or TCM (□). Increased VEGF-A expression was observed after 3 days (▩) and 5 days (▪) of incubation with HIV-TCM as compared with TCM and standard medium.
