FIG. 1.

Recruitment and localization of the N protein to the RTCs and DMVs. (A) Schematic outline of the MHV-N-GFP recombinant virus (not drawn to scale). UTR, untranslated region. (B) LR7 cells inoculated with MHV or MHV-N-GFP were fixed at 6 h p.i. and stained with antibodies directed against nsp2/3 (D4 [24]; kind gift of S. Baker) or nsp8 (anti-p22 antibody [15]; kind gift of M. Denison). Production of newly synthesized viral RNA was visualized by using Click-It detection of RNA. To this end, infected cells were fed with 5-ethynyl uridine from 5.5 to 6.5 h p.i., after which the cells were fixed. (C) HeLa-CEACAM1a cells infected with MHV-N-GFP and control cells (mock) were fixed at 6 h p.i. and processed for immunoelectron microscopy using antibodies against GFP. Arrowheads indicate colocalization sites between N-GFP and either RTC protein markers (B) or DMVs (C).