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. 2017 Nov 27;114(50):E10707–E10716. doi: 10.1073/pnas.1712033114

Fig. 4.

Fig. 4.

BICD2 interacts with HIV-1 capsid in vivo and in vitro. (A) Control and BICD2 knockout HeLa TZM-bl cells were synchronously infected with JRFLg pseudotyped HIV-1 reporter virus (MOI of 0.4). Cells fixed 1 h post infection and PLA assay performed with antibodies to BICD2 and HIV-1 capsid protein p24. (B) Quantification of average fold increase in PLA puncta; 20 or more cells were analyzed in each experiment. Error bar represent the SEM (**P < 0.01). (C) The 293T cells were transfected with HA-tagged WT BICD2 and mutant (BICD2A/V). Cells were lysed 48 h posttransfection and the lysates incubated at room temperature for 1 h with in vitro-assembled HIV-1 CA-NC complexes. Samples were taken either before (INPUT) or after sedimentation through a sucrose cushion (BOUND) and analyzed by WB using anti-HA and anti-p24 antibodies. (D) Binding of BICD2 CC2-CC3 domain. (E) Binding of BICD2 CC2 domain. (F) Binding of BICD2 CC3 domain. A representative experiment is shown.