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. 2015 Sep 14;35(21):2711–2722. doi: 10.1038/onc.2015.332

Figure 1.

Figure 1

Nlp CRBs interfere with apical basal polarity in 3D tissue culture. (a) Localization of overexpressed EGFP-tagged centrosome proteins in 2D culture, as visualized by fluorescence microscopy. (b) Western blots indicate levels of overexpression of EGFP-tagged centrosome proteins after induction (Dox) over time. (c) Top panel: schematic illustrating experimental design for protein induction in 3D culture; bottom panel: DAPI staining of mammospheres, illustrating proficiency or deficiency in basal alignment of nuclei. See also Supplementary Figure S1. (d) Analysis of mammospheres expressing EGFP-Nlp or EGFP-Cep68 by immunofluorescence microscopy. EGFP-Nlp and EGFP-Cep68 are shown in green, Laminin5 and GM130 in red, and DNA in blue (DAPI). See also Supplementary Figure S2. (e) Chimeric mammosphere cluster, illustrating the impact of Nlp CRBs on 3D architecture. Staining as in b. Asterisks indicate parts of the cluster showing glandular morphology and basal laminin5 staining.