FIGURE 8.

Processing of 8E5/LAV envelope glycoprotein and intracellular transport and release of mature virions at 39.5 °C. Actively growing 8E5/LAV cells were divided evenly and incubated at 37 °C with 100 μg/ml IUdR or at 39.5 °C in the presence of the following agents. A, 8E5/LAV culture treated with brefeldin A was assayed for supernatant 8E5/LAV virus and cell lysates for viral gp120 and gp41 by immunoblotting. Results shown are representative of four independent experiments. B, 8E5/LAV culture treated with tunicamycin was assayed for intracellular viral gp120 by immunoblotting. Results shown are representative of four independent experiments. C, 8E5/LAV culture in the presence of cytochalasin D, jasplakinolide, and wortmannin, at the indicated drug concentration. Culture supernatants were assayed for HIV p24 protein, cell lysates were assayed for cell-associated 8E5/LAV RNA copy number by quantitative PCR, and cytotoxicity of the test agents was determined by the MTT assay. Data represent mean ± S.E. (error bars) (n = 3).