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. Author manuscript; available in PMC: 2019 Jun 1.
Published in final edited form as: Nature. 2018 Dec 5;564(7736):444–448. doi: 10.1038/s41586-018-0773-z

Extended Data Figure 8. Purification of the d2d8 stapled ribosome and structural determination by cryo-EM.

Extended Data Figure 8

a, Isolation of the d2d8 stapled ribosome on a sucrose gradient for cryo-EM. Fractions corresponding to the middle of the 70S peak (grey shading) were collected and used in structural studies. The inset shows RNA agarose gel analysis of the stapled ribosome at the different purification stages (30S extract preparation and salt wash) as well as the combined 70S fraction sample. The arrow indicates the position of the stapled rRNA. For source data regarding gels, see Supplementary Fig. 1. The data represent n = 2 independent preparations. b, Fourier shell correlation (FSC) curve, calculated between independent half-maps. The resolution is estimated from the map-to-map correlation at FSC = 0.143 (for a detailed description, see Extended Data Table 1). The electron-microscopy map is coloured according to local resolution. c, Workflow showing the three-dimensional classification and refinement of cryo-EM particles.