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. 2012 Aug 1;23(15):2856–2866. doi: 10.1091/mbc.E10-12-1010

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© 2012 Demelash et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 4: — Silencing hASH1 in pulmonary carcinoid H727 cells leads to reduced migration and invasion. (A) Photomicrographs of immunohistochemical and immunofluorescence staining of hASH1 and p35 expression in H727 cells. Left, nuclear pattern of hASH1 immunoreactivity (top: brown, immunoperoxidase stain,) and cytoplasmic and nuclear distribution of p35 (bottom: green, immunofluorescence, bottom). Right, colocalization of hASH1 (nuclear, granular precipitate in pseudowhite) and p35 (green) in several cells (double staining with immunoperoxidase and immunofluorescence). (B) Phase-contrast photomicrographs of human carcinoid H727 cell migration assay. Down-regulation of hASH1 is associated with reduced cellular migration toward the wound at 24 h. (C) Quantification (bar graph) of the wound areas is expressed as the percentage of the initially bare area at 0 h that is occupied by the migrating cells at 24 h. The reduction in migration associated with shRNA was statistically significant (p < 0.002). (D) Silencing of hASH1 by shRNA significantly blocks invasion in H727 cells (p < 0. 05). The bars represent the means ± SD from three independent experiments.