EIAV replication kinetics and temperature sensitivity assays. (A) Replication analysis of wild-type (WT) EIAV and EIAV with mutations at position d. The procedure employs ED cells which are infected by the virus in sets of three. EIAV replication kinetics is assayed using qPCR, by measuring the virions in the supernatant. (B) Temperature sensitivity measurements; at 41°C, each virus experiences a infection decrease compared to a temperature of 37°C. The same behavior is observed for WT but to a different degree. (C) The infection ratios from 37 to 41°C. The V505T mutation exhibits significant sensitivity with a considerable drop in infection as the temperature is increased. The L512T mutant is also affected in a similar manner, while the other mutants tested show minimal differences. Results are an average of three independent experiments.