ch-TOG and CLASP1 localization during γ-tubulin–independent MT nucleation in the interphase cytoplasm. Oblique illumination fluorescence microscopy of TubG1-mAID-mClover, ch-TOG-mCherry (or CLASP1-mCherry), and SiR-tubulin. (A–D) Three-color images were acquired to show γ-tubulin depletion in the first frame (A and C), followed by two-color imaging every 2 s (B and D). A part of the cytoplasm (boxed in A and C) is magnified to show a nucleation event (B and D). Cells marked in red circles showed no detectable γ-tubulin signals. Arrows indicate MT minus ends.