FIG 5.
Affinity purification of monomeric L-NP from RNP-expressing cells. (A) Transfected cells expressing functional MACV RNP components accumulate eGFP over the course of 72 h posttransfection (hpt). Cells transfected with wild-type (pLWT) or catalytically inactive (pLSDD) L, in combination with pNP and pMG, are shown by fluorescence (top) and bright-field (bottom) microscopy images for each condition at 24, 48, and 72 hpt. (B) Affinity purification of MACV L from the cells shown in panel A at 72 hpt. MACV L was purified from BSR-T7 cells as described in Materials and Methods. Lanes: N, final elution from Ni-NTA purification; F, final elution from anti-Flag antibody-mediated purification. (C) Negative-stain EM images of apo-L purified from insect cells and monodispersed L-NP complexes purified from BSR-T7 cells. Selected regions of the micrographs (white outlined boxes) are shown magnified to the right of the L and L-NP particle images.