Fig 4.

Temsirolimus reduces ANDV release but not viral RNA synthesis. (A) Temsirolimus reduces ANDV release from HMVEC-L cells. HMVEC-L cells were infected with ANDV (MOI of 1), and medium containing temsirolimus or an equivalent amount of the ethanol vehicle control was added to the cells. At 72 h postinfection, the supernatant was removed, and viral titers were determined. Temsirolimus reduced ANDV release by approximately 50% at all concentrations tested. (B) Temsirolimus does not inhibit ANDV RNA production. Total RNA was extracted from HMVEC-L cells treated as described for panel A and subjected to TaqMan analysis of ANDV S segment RNA. Viral transcript levels were normalized to cellular glyceraldehyde-3-phosphate dehydrogenase transcript levels, and data are presented as fold change relative to that of the input virus at 1 h postadsorption. Results were reproduced in two separate experiments performed in quadruplicate. EtOH, ethanol; hpi, hours postinfection.