Basic characterization of TubG1 mAID-mClover cell lines. (A) Immunoblotting of γ-tubulin and α-tubulin for TubG1-mAID-mClover lines (TubG2 intact and KO lines) and the parental line. (B and C) Sucrose gradient centrifugation followed by immunoblotting of γ-tubulin for the parent line (B) and the TubG1-mAID-mClover line (C). (D) Spindle dynamics in TubG1-depleted cells (TubG2 KO background) and control cells. Time 0 corresponds to nuclear envelope breakdown (NEBD). (E) Mitotic duration (NEBD to anaphase onset) of γ-tubulin–depleted cells. Ana, anaphase; Dox, doxycycline; Int, interphase; Meta, metaphase; Telo, telophase; Cyto, cytokinesis.