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Figure 6 | Retrovirology

Figure 6

From: TNPO3 protects HIV-1 replication from CPSF6-mediated capsid stabilization in the host cell cytoplasm

Figure 6

TNPO3 KD inhibits HIV-1 replication by shifting CPSF6 to the cytoplasm. (A) Expression level of CPSF6 and TNPO3 in TZM-bl cells stably transduced with control or CPSF6 KD vectors and transfected with scrambled or TNPO3 specific small interfering RNA (siRNA). Cell lysate was probed in western blots with anti-TNPO3 antibody (upper panel), anti-CPSF6 antibody (middle panel) and anti-β-actin antibody (lower panel). (B) Immuno-fluorescence localization of endogenous CPSF6 (green) in control (Ctrl) KD or TNPO3 KD TZM-bl cells. DAPI staining (blue) was used to mark the nuclear compartment. (C) Cell fractionation to identify the cellular localization of endogenous CPSF6. Expression of tubulin in the cytoplasm and histone 3 (H3) in the nucleus was assessed to verify the fractionation. (D) Stable ctrl KD and CPSF6 KD cells transfected with scrambled or TNPO3 siRNA were challenged with WT or CA mutant HIV-1NL4-3GFP reporter viruses. After 72 hours GFP expression was checked by flow cytometry. The fold inhibition of infectivity due to TNPO3 KD is shown. Data represent one of at least three independent experiments. Error bars represent ± SEM (n = 3).

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