FIGURE 2.

Src localizes to or close to mitotic spindles. A, HeLa S3 cells inducibly overexpressing c-Src were fixed in 4% paraformaldehyde for 3 min and extracted with 0.5% Triton X-100 for 5 min at room temperature, followed by further fixation in 4% paraformaldehyde for 15 min. Fixed cells were stained for c-Src (red), α-tubulin (green), and DNA (blue) (panels a–c). Cells were stained with either anti-α-tubulin antibody (panel d) or anti-Src antibody (panel e) alone. B, HeLa S3 cells expressing c-Src were treated with 20 μm PP2 and then fixed and stained for c-Src, α-tubulin, and DNA as described for A. C, c-Src-inducible HeLa S3 cells were treated with or without 1 μg/ml doxycycline (Dox) and then fixed as described for A. Fixed cells were stained for anti-Src phospho-Tyr⁴¹⁸ antibody (red), α-tubulin (green), and DNA (blue). D, HeLa S3 cells overexpressing c-Src were treated with 20 μm PP2 for 3 h, fixed, and stained as described for C. Scale bars = 10 μm.