Fig. 2: PORT-HS-AFM and cryo-EM uncover impact of monobodies on CrSAS-6 ring assembly and stacking.
From: Tuning SAS-6 architecture with monobodies impairs distinct steps of centriole assembly
a-d PORT-HS-AFM equilibrium analysis of CrSAS-6_NL alone (a), or together with MBCRS6-1 (b), MBCRS6-13 (c), or MBCRS6-15 (d), with higher magnification views on the right showing in each case two representative examples of assemblies. Scale bars: 25 nm; z scale color bar, –0.5 to 10 nm. All measurements were performed at least twice, with at least 5 field of view imaged in each experiment. e-h Transmission cryo-EM of ring stacking assay42 and corresponding symmetry distributions based on particle classification of CrSAS-6_NL alone (e, f) or CrSAS-6_NL and a 50% molar excess of MBCRS6-1 (g, h). Note that spokes can be difficult to discern and be more numerous than 9, due to partially out of register stacking of SAS-6 ring polymers. The orange arrows point to protrusions observed on the outside of SAS-6 rings, corresponding to the density contributed by the monobody. Data represent mean ± SD. Particles numbers from at least 3 independent experiments: CrSAS-6_NL N = 2612, CrSAS-6_NL + MBCRS6-1 N = 3101.
