Fig. 1

eEF1A binds to HIV-1 genomic RNA. a TMZ-bl cells were incubated with HIV-1 for 2 h at 4 °C and 2 h at 37 °C in the presence or absence of nevirapine followed by RC-co-IP using anti-eEF1A or anti-eIF3A antibodies as indicated. The level of RNA extracted from IP product was measured by RT-PCR targeting HIV-1 5’UTR. Data are presented as means ± SD from 3 independent experiments. TMZ-bl cells were also transfected with biotin-labeled RNAs derived from HIV-1 genomic 5’UTR, HIV-1 RT and luciferase sequences followed by RC-co-IP. The RNAs in cell lysates and recovered from IP using anti-biotin antibody (b, upper panel) or anti-eEF1A antibody (b, lower panel) were detected by dot blot using streptavidin-peroxidase. The blot signals were quantified using ImageQuant programme and the relative PI unit of IP product to lysate product are presented in c. The biotin-labelled d 5’UTR, e RT or f luciferase RNAs were immobilized on the biosensors coupling with streptavidin. The associations of RNAs with eEF1A were measured with OctetRed system using 90nM or 30nM purified eEF1A protein and referenced using 90 nM of BSA in kinetic buffer. The dissociation was measured by moving the biosensor to wells containing kinetic buffer only. g The association and dissociation of immobilized 5’UTR, RT and luciferase RNA with 90 nM of eEF1G was also measured as a control. Data are representative of 3 independent experiments. * indicates p < 0.05