Figure 5.

Cryo-ET of nodavirus RNA replication complexes in mammalian cells. (A,F) Indistinguishable spherules are induced on the outer mitochondrial membrane in mammalian BSRT7/5 cells transfected with either wildtype nodavirus RNA1 (A) or with RNA1-noB2 (F). Arrowheads: black = outer mitochondrial membrane; white = inner mitochondrial membrane; red = spherules, filled with dsRNA; yellow = crown-like structures at the spherule open necked connections to the cytoplasm. (B,G) Close-up side views of spherules with crowns visible in the plane of sectioning. (C,H) Close-up top views of crowns. (D,I) Cross sections of EM density maps of subtomogram-averaged side views of crowns. Arrowheads: light blue = apical lobe; dark blue = basal lobe; green = leg; orange = floor. (E,J) Cross sections of EM density maps of subtomogram-averaged top views of crowns with blue arrowheads pointing to the central crown turret and green arrowheads to the extended legs. Tomogram movies corresponding to panels A and F and Fourier Shell Correlation (FSC) curves corresponding to panels (D,E,I,J) are provided in Supplementary Movies S1 and S2 and Supplementary Figure S1.