Figure 1
From: Ebola virus VP24 interacts with NP to facilitate nucleocapsid assembly and genome packaging
NP, VP35, and VP24 interact independently with each other. (a–c) HEK 293 cells were co-transfected with pCAGGS-NP-FH and pCAGGS-Myc-VP35 (a), pCAGGS-NP-FH and pCAGGS-VP24 (b), or pCAGGS-Myc-VP35 and pCAGGS-VP24 (c). Lysates were immunoprecipitated with mouse anti-FLAG (a,b), mouse anti-Myc (c), or isotype control (Iso; a,b) antibodies, and immunoprecipitation (IP) and whole cell lysate (WCL) fractions were subjected to Western blot (WB) with mouse anti-FLAG, mouse anti-Myc, rabbit anti-VP24, and rabbit anti-β-tubulin antibodies. The precipitated proteins in the IP fraction are labeled with arrowheads, and the light chain (LC) of the mouse anti-Myc antibody is indicated with an arrow (a,b). IP/Western blot data are representative of at least three independent experiments. Western blots were cropped for presentation; full-length blots are available in Supplementary Fig. 6. (d–f) Hela cells were co-transfected with pCAGGS-NP-FH and pCAGGS-Myc-VP35 (d), pCAGGS-NP-FH and pCAGGS-VP24 (e), or pCAGGS-Myc-VP35 and pCAGGS-VP24 (f). Following fixation and permeabilization, cells were stained with rabbit anti-FLAG (d), mouse anti-FLAG (e), mouse anti-Myc (d,f), and/or rabbit anti-VP24 (e,f) antibodies prior to staining with AlexaFluor 488 goat anti-mouse and AlexaFluor 546 goat anti-rabbit secondary antibodies. Coverslips were mounted with ProLong Gold Antifade Mountant containing DAPI to visualize the nuclei. White scale bars represent 10 uM.
