Supplemental Material

This article contains the following supporting material:

• Figure 1 - (A) The EGFP-tubulin mutants are expressed in equivalent amounts and at their expected molecular weight of 81 kDa when transfected in HeLa cells. Adherent HeLa cells were either non transfected (n.t.) or transfected with EGFP-beta tubulin^WT, EGFP-beta tubulin^S172A, EGFP-beta tubulin^S172E or EGFP-beta tubulin^S172D, before extraction in Laemmli buffer. Equal volumes of total cell extracts were analyzed by Western blotting with GFP Ab. (B) EGFP-beta tubulin^S172D binds to alpha tubulin in cells. HeLa S3 cells were either non transfected (n.t.) or transfected with EGFP-beta tubulin^S172D (S172D) before cell lysis and immuno-precipitation with anti-alpha tubulin YL1/2 mAb. Immuno-precipitates were submitted to SDS PAGE to show amounts of total tubulin (top), and to Western blotting with GFP Ab to detect EGFP-beta tubulin (bottom).