Whole-cell protein extracts were prepared in cold cell lysis buffer [50 mM tris-HCl (pH 7.5), 150 mM NaCl, 0.1% sodium dodecyl sulfate, 1% Triton X-100, 0.5% sodium deoxycholate, and 5 mM EDTA] supplemented with protease and phosphatase inhibitors. Proteins were quantified using BCA protein assay, resolved on SDS–polyacrylamide gel electrophoresis gels (4 to 20%), and transferred to nitrocellulose membranes. Alternatively, cells were counted and directly resuspended in Laemmli buffer (1×). Protein samples were normalized to either tubulin or β-actin. A detailed list of antibodies used in the study can be found in table S10.

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