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. 2015 Aug 5;6:7929. doi: 10.1038/ncomms8929

Figure 6. Phosphomutant Mto2[24A]-GFP forms interphase-like puncta in mitosis.

Figure 6

(a) Localization of wild-type and phosphomutant Mto2-GFP, as indicated, in mitotic cells. Note robust puncta of Mto2[24A]-GFP associated with the nuclear envelope (NE) relative to other phosphomutant proteins, and absence of puncta of wild-type Mto2-GFP (except for spindle pole bodies; SPBs). (b) Quantification of total fluorescence signal of wild-type and phosphomutant Mto2-GFP at the NE in mitotic cells (n=11, 24, 12, 30, 16, 13 and 12 cells, respectively). Fluorescence signal at SPBs was excluded from analysis (see Methods). Median with interquartile range is shown in red. ***P<1e-4 (Wilcoxon rank-sum test). (c) Localization of wild-type and phosphomutant Mto2-GFP, as indicated, in mitotic mto1[NE] cells, together with SPB marker Cut12 fused to tandem-dimer Tomato (Cut12-tdT). Because Mto1/2 complex in mto1[NE] cells does not localize to SPBs, this demonstrates that NE-associated puncta of Mto2[24A]-GFP and Mto2[17A]-GFP form independently of localization to the SPB. All images are Z-projections; individual Z-sections show NE-associated Mto1/2 complex on the NE and not in nuclear interior (see Supplementary Fig. 6). Scale bars, 5 μm.