Figure 1.

INI1 is dispensable for HIV-1 transduction. a. Inhibition of ini1 expression by shRNA-producing lentiviral vector. Ethidium bromide-stained agarose gel analysis of RT-PCR amplification products of cytoplasmic RNA from INI1 knockdown P4.2 cells (INI1i) as well as in P4.2 cells transduced with a control (C) lentiviral vector. Cytoplasmic RNA was obtained using RNeasy kit (Qiagen) and RT-PCR was performed using SuperScript one-step RT-PCR with Platinum Taq kit (Invitrogen) with following primers: ini1, 5'-TCTGGAGGCGACTAGCCACTGTG-3' (forward primer) and 5'-GATCACAGCTGGGTCATGGTCATC-3' (reverse primer); beta-actin, 5'-TGACGGGGTCACCCACACTGTGCCCATCTA-3' (forward primer) and 5'-CTAGAAGCATTTGCGGTGGACGATGGAGGC-3' (reverse primer). b. Transduction efficiency in the same cells was determined by GFP fluorescence-activated cell sorter (FACS) analysis with a FACStrak apparatus (Becton Dickinson) 2 days after exposure to VSV-G-pseudotyped GFP-expressing HIV-1-derived lentiviral vector at the indicated MOI. Experiments were done in duplicate and columns represent the mean percentage of transduced cells, with mean fluorescence intensity (MFI) indicated below. c. Reduced HIV-1 replication in INI1 knockdown cells. INI1 knockdown P4.2 cells were infected with HIV-1 at a MOI of 0.5. HIV-1 replication was assayed by p24 ELISA in the culture supernatant 6 days later.