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. 2015 Mar 2;208(5):613–627. doi: 10.1083/jcb.201408026

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© 2015 Chung et al.

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Figure 6. — K17 and hnRNP K are required for the secretion of factor(s) necessary for tumor cell growth. (A) Anchorage-independent growth assay using A431 cells stably expressing SCR or KRT17 shRNA in soft agar. Cells were grown in conditioned medium from A431 SCR or KRT17 shRNA cells. Bar, 1 cm. (B) Colony numbers of SCR shRNA cells grown in conditioned medium from SCR or KRT17 shRNA cells from A were quantitated using ImageJ and normalized to those grown in conditioned medium from A431 SCR shRNA cells. Data from nine experimental repeats (each performed in triplicate) are represented as mean ± SEM (error bars). *, P < 0.05. (C) Matrigel invasion assay of A431 SCR shRNA cells. Cells were allowed to invade using conditioned medium from TPA- or DMSO-treated A431 SCR or KRT17 shRNA cells as a chemoattractant for 48 h. Nuclei of invaded cells were stained with propidium iodide and imaged under a fluorescent microscope. Bar, 100 µm. (D) The number of invaded cells from C was quantitated using ImageJ and normalized to the cells exposed to conditioned medium from DMSO-treated A431 SCR shRNA cells. Data from three experimental repeats (each performed in triplicate) are represented as mean ± SEM (error bars). *, P < 0.01. (E) Anchorage-independent growth assay using A431 SCR shRNA cells in soft agar. Cells were grown in conditioned medium from A431 SCR shRNA cells transfected with NS (NS siRNA cells) or hnRNP K (RPK1; hnRNP K siRNA cells) siRNA. Bar, 1 cm. (F) Colony numbers from E were quantitated using ImageJ and normalized to cells grown in conditioned medium from cells expressing NS siRNA. Data from three experimental repeats (each performed in triplicate) are represented as mean ± SEM. *, P < 0.005.