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. 2020 Mar 24;219(5):e201905018. doi: 10.1083/jcb.201905018

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© 2020 Auckland et al.

This article is available under a Creative Commons License (Attribution 4.0 International, as described at https://creativecommons.org/licenses/by/4.0/).

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Figure S1. — CENP-F microtuuble binding mutant rescues. (a) Immunofluorescence microscopy images of the CENP-E rescue experiment with CENP-F MTBD mutants. HeLa-K cells were treated with control or CENP-F siRNA and rescued with an empty vector, CENP-F^mEmerald, CENP-F^mEmeraldΔnMTBD, CENP-F^mEmeraldΔcMTBD, or CENP-F^mEmeraldΔn+cMTBD before being stained with DAPI and antibodies against CENP-C and CENP-E. Scale bar, 5 µm. (b) Immunofluorescence microscopy images of the K-K distance rescue experiment with CENP-F MTBD mutants. HeLa-K cells were treated with control or CENP-F siRNA and rescued with an empty vector, CENP-F^mEmerald, CENP-F^mEmeraldΔnMTBD, CENP-F^ΔcMTBD, or CENP-F^GFPΔn+cMTBD before being stained with DAPI and an antibody against CENP-C. Scale bar, 5 µm. (c) Immunofluorescence microscopy images of the cold-stable rescue experiment with CENP-F MTBD mutants. HeLa-K cells were treated with control or CENP-F siRNA and rescued with an empty vector, CENP-F^mEmerald, CENP-F^GFPΔnMTBD, CENP-F^GFPΔcMTBD, or CENP-F^GFPΔn+cMTBD before being incubated on ice for 10 min and stained with DAPI and antibodies against CENPC and α-tubulin. Scale bar, 5 µm.