γ-tubulin ^ΔGTP+KD cells form disrupted spindles but display non-centrosomal microtubule formation. (A) Analysis of the mean mitotic index. n = >2,000 cells per condition from N = 3 independent experiments. (B) The mean percentage of mitotic cells that displayed disrupted spindles. n = >200 cells per condition from N = 3 independent experiments. (C and D) Images of fixed mitotic γ-tubulin^KD, γ-tubulin^ΔGTP+KD, and γ-tubulin^WT+KD cells. Single-channel images (maximum-intensity projections) and overlays show γ-tubulin (immunofluorescence; C) or GFP (fluorescent signal; D; magenta), α-tubulin (green), and DNA (blue). (E) Quantification of the mean percentage of mitotic cells that display >2 microtubule foci at 5 min after nocodazole washout. n = >45 cells per condition from N ≥ 3 independent experiments. (F and G) Images of fixed mitotic γ-tubulin^KD and γ-tubulin^ΔGTP+KD cells 5 min after nocodazole washout. Single-channel images (maximum-intensity projections) and overlays show γ-tubulin (immunofluorescence; C) or GFP (fluorescent signal; D; magenta), α-tubulin (green), and DNA (blue). Scale bars = 5 µm. Error bars = standard deviation (A, B, and E).