DEAR1 is a negative regulator of TGF-β–induced migration and EMT

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DEAR1 is a negative regulator of TGF-β–induced migration and EMT DEAR1 is a negative regulator of TGF-β–induced migration and EMT. A, phase contrast images of DshR and CshR clones with or without TGF-β (2 ng/mL) in 3D culture at indicated times. DEAR1 is a negative regulator of TGF-β–induced migration and EMT. A, phase contrast images of DshR and CshR clones with or without TGF-β (2 ng/mL) in 3D culture at indicated times. Experiments were repeated twice with similar data obtained. B, Western analysis of DshR and CshR clones with or without TGF-β in 3D culture. Cells collected at time points indicated lysed in 1× SDS sample buffer. C, phase contrast images of DshR clones (DshR1-T and DshR2-T) and CshR clones (CshR1-T and CshR2-T) with or without (data not shown) TGF-β (2 ng/mL) in 3D culture for 5 days. Red arrows trace movement of TGF-β–treated DshR clones through the matrix. D, cell migration distance at each time point comparing 76N-E6–DEAR1-KD clones with or without TGF-β treatment (DshR-T vs. DshR-C) and control clones with or without TGF-β treatment (CshR-T vs. CshR-C). The values shown are means of 150 to 200 cells. E, wound-healing assay of DEAR1-KD (DshR1 and DshR2) and control clones (CshR1 and CshR2) of 76N-E6 (left) and MCF10A cells (DshR and CshR, right) with or without TGF-β treatment (2 ng/mL) for 24 hours. F, Western analysis of EMT markers in 76N-E6–DEAR1-KD (DshR1 and DshR2) and control clones (CshR1 and CshR2) with or without TGF-β treatment (4 ng/mL) for 4 days. Nanyue Chen et al. Cancer Discovery 2013;3:1172-1189 ©2013 by American Association for Cancer Research