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. 2014 Apr 14;24(8):896–903. doi: 10.1016/j.cub.2014.03.006

Figure 3.

Figure 3

Assembly of Multimeric Puncta In Vivo

(A) Images showing puncta of Mto1[NE]-GFP, Mto1[bonsai]-GFP, and Mto1[9A1-bonsai]-GFP in wild-type backgrounds (mto2+) and absence of puncta in mto2Δ backgrounds. Mto1[9A1-bonsai]-GFP puncta diffuse rapidly but are prominent in movies (see Movie S3). Full-length Mto1-GFP displays a weak but still detectable signal in mto2Δ backgrounds, due to its intrinsic ability to enrich at many MTOC sites ([9]; see also Figure 1A).

(B) Mto2-GFP puncta together with mCherry-tubulin (mCh-Atb2) in mto1[bonsai] and mto1Δ cells. Arrows indicate examples of puncta in mto1Δ cells, which are relatively faint and seen more easily in movies or when Mto2-GFP is overexpressed (see Movie S5 and Figure S3E). Wild-type cells (mto1+ mto2+) provide a negative control for GFP fluorescence.

(C) Mto2-13myc is coimmunoprecipitated with 6His-FLAG-GFP-Mto2 (HFG-Mto2), in both mto1+ and mto1Δ backgrounds. Western blots were probed with antibodies against GFP, Myc, and Mto1. Scale bars, 5 μm.