Extended Data Fig. 2: Data acquisition and processing of the pre-cleavage complex.
From: Mechanism of co-transcriptional cap snatching by influenza polymerase
a, Fluorescence scan of the native PAGEs analyzing GraFix gradient fractions of the pre-cleavage complex preparation (n = 1). The image is an overlay of Cy5 and ATTO532 signals. In magenta is the scan for the Cy5 channel of the labeled template DNA, and in green is the scan for the ATTO532 channel of the labeled 3′ vRNA. Highlighted fractions were combined and used for cryo-EM analysis. b, Flowchart illustrating the key steps of the processing pipeline for obtaining the structure of the pre-cleavage complex. c, Consensus density map of the pre-cleavage FluPol-Pol II-DSIF EC complex colored by underlying protein components. d,e, Masks used for focused refinements and their resulting maps. f, Gold-Standard Fourier shell correlation plots of the consensus and focused maps. g, Local resolution of consensus and focus refinement maps. Local resolution estimations were performed in RELION. h, Angular distribution of the consensus refinement and focused refinements plotted with Warp.
