Fig. 3
From: Alpha-enolase in viral target cells suppresses the human immunodeficiency virus type 1 integration
Effects of ENO1 overexpression in HIV-1 target cells on postentry step. a Confirmation of ENO1-V5 expression in TZM-bl cells. Endogenous ENO1 was detected with an anti-ENO1 antibody and exogenous ENO1 (ENO1-V5) was detected with an anti-V5 antibody. b HIV-1LAV-1 infectivity in ENO1-V5 expression vector-treated TZM-bl cells. The infectivity was examined at 2 days postinfection with HIV-1LAV-1 and assessed on the basis of the luciferase activity in lysates of vector-treated TZM-bl cells. The infectivity in the control-vector-treated TZM-bl cells was set as 100%. c Flow cytometry monitoring of HIV-1 receptor CD4 and coreceptors CCR5 and CXCR4. Cells stained with the isotype control antibody are indicated as negative control (black line). The expression levels of CD4, CCR5 and CXCR4 in control-vector- (red line) or ENO1-V5-expression vector (blue line)-treated TZM-bl cells are shown. Each protein was detected on the infection day. d Effect of ENO1-V5 expression in TZM-bl cells on viral reverse transcription. The amount of R/gag products of viral reverse transcription was determined by quantitative real-time PCR analysis. e Effect of ENO1-V5 expression in TZM-bl cells on viral cDNA nuclear import. The amount of 2-LTR circle products was determined by quantitative real-time PCR analysis. f Integration efficiency of HIV-1LAV-1 in either control- or ENO1-V5-expression vector-treated TZM-bl cells. Relative amount of Alu-gag products was determined by nested-PCR. The integration efficiency in the control-vector-treated TZM-bl cells was set as 100%. Data are mean values ± SE from triplicate tests. The significance of difference (Student’s t-test) is indicated as follows: **, p < 0.01; *, p < 0.05; n.s., not significant