Associations of Tks5 WT and mutants with microtubules in MDA-MB-231, MCF7, and SK-BR-3. (A) Immunofluorescence staining of α-tubulin and endogenous Tks5 in methanol-fixed MDA-MB-231 cells. The arrowheads indicate the regions of colocalization between TKS5 and α-tubulin. (B) Immunofluorescence staining of α-tubulin and transiently expressed GFP-Tks5 in methanol-fixed MDA-MB-231, MCF7, and SK-BR-3 cells. (C) Captured images of GFP-Tks5 and mCherry-α-tubulin in a live MDA-MB-231 cell from Video 3. (D–F) Immunofluorescence staining of α-tubulin and transiently expressed sequential (D) and internal (E) deletion and point (F) mutants of GFP-Tks5 in PFA- or methanol-fixed MDA-MB-231 cells. (G) Schematic drawing of Tks5 point mutants and the summary of the mutants’ localization and microtubule bundling. (H) Immunofluorescence staining of α-tubulin and transiently expressed GFP-Tks5 aa 363–1118 in methanol-fixed MDA-MB-231, MCF7, and SK-BR-3 cells. (A–F and H) Scale bars, 10 μm.