mTOR activity is not necessary for Andes virus glycoprotein-mediated entry. (A) To confirm pseudoparticle specificity, virions were neutralized with antibodies specific for the ANDV or the VSV glycoprotein. Virions were adsorbed to HMVEC-L cells (MOI of 14) for 6 h, and the inoculum was removed and replaced with fresh medium. Luciferase expression was measured at 3 days postinfection. RLU, relative light units. (B) Temsirolimus did not block ANDV or VSV pseudoparticle entry into HMVEC-L cells. Cells were pretreated for 1 h with temsirolimus or an equivalent amount of ethanol and infected for 6 h with pseudoparticles in drug-containing medium, and the inoculum was removed and replaced with fresh medium lacking temsirolimus and ethanol. Luciferase expression was measured at 3 days postinfection. Results were reproduced in two separate experiments performed in triplicate. Env, envelope protein; Ab, antibody.