Mutant CAs are sensitive to REAF restriction. (A) Infection of HeLa-CD4-shREAF cells with HIV-1NL4.3 CA N74D (MCR) renders it susceptible to REAF compared to the viral pseudotype with wild-type CA (HIV-1NL4.3 [MCR]) (P < 0.001). (B to E) Titers of HIV-1NL4.3 (MCR), HIV-1NL4.3 CA P38A (MCR), HIV-1NL4.3 CA G89V (MCR), and HIV-1NL4.3 CA G94D (MCR) following a challenge of HeLa-CD4 and HeLa-CD4-shREAF cells. (F) Fold changes in HIV-1NL4.3 CA P38A (MCR), HIV-1NL4.3 CA G89V (MCR), and HIV-1NL4.3 CA G94D (MCR) on HeLa-CD4-shREAF cells compared to wild-type CA (HIV-1NL4.3 [MCR]) show that they are also susceptible to REAF (all P < 0.001 [*]).