Fig. 6
ABIN1 specifically decreases the ubiquitination of Tat at Lys71. a HEK-293T cells were treated with siRNAs and plasmids similar as in Fig. 4a as indicated, except that Flag-Tat-K71R mutant was used to assess the ubiquitination of wild-type Tat and TatK71R mutant in the presence or absence of ABIN1. b HeLa cells were infected with HIV-1(Luc) or HIV-1(Luc-K71R) at the same MOI of 0.2, replication activity (Repl. activity) was assessed by luciferase assays 48 hpi to evaluate the importance of Lys71 in Tat. c, d HEK-293T cells were treated with siRNAs targeting ABIN1 (siABIN1-1, siABIN1-2) or siNC (c) or transfected with wild-type ABIN1 or QE2 mutant encoding vectors and challenged with HIV-1(Luc) or HIV-1(Luc-K71R) (d), then subjected to luciferase assays 24 hpi to monitor the replication efficiency, and the cell lysates were collected and cleared for detection of knockdown efficiency and overexpression level of ABIN1/mutant. The blots are representatives of at least three independent experiments achieving similar results