Extended Data Fig. 7: Cryo-EM analysis and image processing workflow for hexameric SenDRT9 RT-ncRNA complex.
From: Protein-primed homopolymer synthesis by an antiviral reverse transcriptase
a, Representative micrograph from the SenDRT9 RT-ncRNA hexameric complex, revealing a monolayer of evenly distributed particles. b, Initial reconstruction from on-the-fly image analysis that was used to generate de novo templates used for particle picking. c, Representative 2D Classes from 6,011,113 template-picked particles. d, Initial volumes from a 7-class ab initio reconstruction of 1,411,369 particles. Selected volumes for downstream processing are shown with a green background. e, Final steps used to sort selected particles in d to yield the final reconstruction. f, Final (D3) reconstruction of the SenDRT9 RT-ncRNA hexamer that refined to 2.6 Å and was used for model building. The map is shown as a partially transparent surface, with RT subunits shown in blue/cyan and ncRNA subunits shown in orange/salmon. g, Plot of Cryosparc’s gold standard Fourier shell correlation for the map in f. h, Plot of particle view distribution reveals mild anisotropy but no missing views.
