Figure 3.

The MGM Holocomplex Is a Potent Microtubule Nucleator In Vitro

(A) DAPI fluorescence assay for microtubule (MT) polymerization, using porcine brain tubulin and the indicated complexes and concentrations. The MGM holocomplex nucleates MTs in a dose-dependent manner. The estimated molarity of MGM is based on assumption that Mto1/2[bonsai] and γ-TuSC^Alp6-MBP:Mzt1 coassemble into a structure similar to the mammalian γ-TuRC (see STAR Methods). Additional controls are shown in Figures S3A–S3C.

(B) Rhodamine-tubulin fluorescence microscopy assay for MT polymerization.

(C) Quantification of MT polymerization for experiments in (B). Each data point represents total MT fluorescence within a randomly chosen field. Inset shows expanded scale for non-MGM samples. Quantification of MT number per field is shown in Figure S3D. Scale bar, 10 μm.

See also Figure S3.