Proteins used in the TIRF experiments. (A) Schematic representation of the proteins with color code as in Fig. 5. VASH1 (V1) is represented in blue, VASH2 (V2) in orange, SVBP in purple, sfGFP in green, tags (myc, FLAG, His) in gray. Vertical bars, catalytic triads. FL, full length; CD, core domain; Nt, N-terminal region; Ct, C-terminal region. For most proteins, an inactive protein (dead version, with the catalytic cysteine residue mutated into an alanine residue) was also used. (B) SDS-PAGE and immunoblots of a complete set of VASH1–SVBP protein preparations. Uncropped images are shown in source data. (C) Immunoblot of three separate brains tubulin preparations (1, 2, 3), and of tyrosinated- and detyrosinated-HeLa tubulin preparations (Tyr, deTyr). These samples were co-analyzed with extracts from HEK293T cells transfected with various mCherry α-tubulin variants (Tyr, deTyr, ∆2). Uncropped images are shown in source data. The mean content of the different forms of α-tubulin present in a brain tubulin preparation was estimated after normalization to total α-tubulin levels (and antibody sensitivity (as in Aillaud et al., 2016) to be of 43, 49.5, and 7.5%, for tyrosinated-, detyrosinated-, and ∆2-tubulin respectively. HeLa tubulin was either 100% tyrosinated or 100% detyrosinated. Taxol-stabilized Tyr-MTs and deTyr-MTs were composed of 65% of HeLa tubulin and 35% of brain tubulin (see Materials and methods). Source data are available for this figure: SourceData FS1.