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Fig. 2 | Journal of Otolaryngology - Head & Neck Surgery

Fig. 2

From: Introduction and expression of PIK3CAE545K in a papillary thyroid cancer BRAFV600E cell line leads to a dedifferentiated aggressive phenotype

Fig. 2

Expression of PIK3CAE545K in a BRAF-mutant PTC cell line promoted colony formation and cell migration in vitro. A Clonogenic assay of LAM1EV and LAM1:PIK3CAE545K cells seeded into six-well tissue culture plates at a density of 1,000 cells per well. After 10 days, cells were fixed and stained with 0.5% crystal violet. Brightfield microscopy was used to quantify colony formation, with a colony defined as being ≥ 50 cells. For quantification, three representative fields per well were counted for the number of colonies and this was completed for both the parental and mutant cell lines. B Cell migration was quantified using fluorometric analysis. Cells were seeded into the top chamber containing serum-free media at a density of 5 × 104 cells per chamber and each chamber was then placed in a well containing media with serum. Cells which migrated through to serum-containing media after 24 h were quantified using fluorometric detection as described using the standard protocol provided. Fluorescence readings were completed using a microplate reader and representative images were collected using brightfield microscopy

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