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A
Schematic representation of EB1 constructs and domain functions. CLIP170 (lower) uses CAP‐Gly domains to interact with EB1's EEY motif, and secondary SxIP‐mediated interactions with EB1's EBH domain.
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B
Representative IF images in CHME3 stably expressing Flag alone control or the Flag‐tagged EB1 constructs depicted in (A) co‐stained for Tyrosinated‐MTs (Tyr‐MTs), and the nucleus (Hoechst). Scale bar, 10 μm.
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C
HIV‐1-VSV‐luc infection is enhanced in CHME3 depleted of endogenous EB1 and stably expressing full‐length (FL) EB1, but not in control Flag or any of the other EB1 constructs depicted in (A).
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D
Increasing amount of FL‐EB1, but not ΔEEY results in a dose‐dependent enhancement of HIV-1‐VSV-luc infection in transiently transfected CHME3 cells.
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E, F
HIV-1‐VSV-luc (E) or HIV‐1-VSV‐ZsGreen at various dilutions (corresponding to multiplicities of infection (m.o.i) 0.06, 0.03, and 0.02), (F) infection is reduced in differentiated THP‐1 depleted of CLIP170 determined by luciferase assays or FACS analysis, respectively.
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G
HIV‐1-WT-luc infection is decreased in CLIP170‐depleted CHME3 cells.
Data information: Data in C‐G are mean values from three independent experiments ± SEM. Statistical significance was determined by one‐way ANOVA (C, D) or
‐test (E‐G). *
< 0.001. Molecular weight markers (in kDa) are shown to the right of WBs.
