Fig. 5 | Nature Communications

Fig. 5

From: Hexameric NuMA:LGN structures promote multivalent interactions required for planar epithelial divisions

Fig. 5

NuMA:LGN cortical protein networks are essential to promote spindle orientation. a Schematic representation of GFP-NuMA constructs decoupling NuMA cortical recruitment and NuMA:LGN oligomerization. b Confocal xy (left) or xz (right) sections of mitotic HeLa cells expressing the indicated GFP-NuMA or GFP-NuMA-GoLoco constructs, treated with DMSO or MLN8237. Cells were stained with DAPI (blue). Scale bars, 5 μm. c Dot-plot illustrating the distribution of the mitotic spindle angles of metaphase HeLa cells shown in panel b. For each condition, means ± SEM are shown for three independent experiments, with n > 56. ****p < 0.0001; ***p < 0.001 and *p < 0.05 by Krustal-Wallis test. d Confocal xz sections of metaphase HeLa cells depleted of endogenous NuMA and transfected with mCherry-NuMA wild-type or mCherry-NuMA-ΔMT. Cells were stained with γ-tubulin (yellow) and DAPI (cyan). Scale bars, 5 μm. e Dot-plot illustrating the distribution of the spindle axis angles for HeLa cells visualized in panel d, with control cell lines expressing a scrambled shRNA or the shRNA-NuMA used to deplete the endogenous protein. For each condition, means ± SEM are shown for four independent experiments, with n > 55. ****p < 0.0001 by Krustal-Wallis test. f Model of the cortical dynein/dynactin:NuMA:LGN:Gαi protein network pulling on astral MTs based on our studies

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