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. 2017 Apr 3;216(4):961–981. doi: 10.1083/jcb.201611060

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© 2017 Mosalaganti et al.

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Figure 4. — Validation of the RZZ model. (A) Elution profiles and SDS-PAGE of SEC experiments on Zwilch, ZW10, or their stoichiometric combination. Molecular mass is indicated in kilodaltons. AU, arbitrary units. (B) Top: 3D reconstruction of the negatively stained ROD^1–1,250ZZ complex. Bottom: the density corresponding to a model of the ROD^1–1,250ZZ complex was cut out of the complete RZZ complex for comparison. (C) NiNTA-gold labeling of the N-terminal histidine tag of ROD. The bottom-right panel represents part of a field in which labeling of ROD was performed after proteolytic removal of the His tag. (D) Micrographs of negatively stained GFP-RZZ or ROD–Zwilch–GFP-ZW10 complexes. Bars, 10 nm. (E) Summary of positions identified in experiments in C and D mapped onto the molecular model of RZZ.