the substrates used in this study, with stiffness values of 0.2, 2, 16, and 64 kpa  (Advanced Biomatrix Inc)

Journal: Oncogene

Article Title: Mediator kinase CDK8/CDK19 drives YAP1-dependent BMP4-induced EMT in cancer

doi: 10.1038/s41388-018-0316-y

Figure Lengend Snippet: BMP4-induced EMT is suppressed on compliant substrates. a Phase contrast images of cells plated on soft (0.5 kPa) and rigid (8 kPa) fibronectin-coated hydrogels and treated with 10 nM BMP4 for up to 5 days to monitor any changes in morphology (Methods). b Immunofluorescence images of BMP4-treated OvCa429 and Panc1 cells plated on soft (0.5 kPa) and rigid (8 kPa) fibronectin-coated hydrogels as described in Methods. Cells were then immunostained for E-cadherin and phalloidin/actin as indicated. Scale bars = 20 μm. c Realtime quantitative PCR of SNAI1 and SNAI2 in OvCa429, Panc1, and Py2T cells plated on soft (0.5 kPa) and rigid (8 kPa) hydrogels followed by treatment with 10 nM BMP4 (Methods). Values are normalized to their untreated control. Graphs are representative of two independent biological trials done in triplicate. *p < 0.05, **p < 0.01, ***p < 0.001

Article Snippet: For rigidity experiments, cells were plated on 0.5 kPa (soft) or 8 kPa (rigid) substrates obtained from Matrigen Life Technologies (Brea, CA, USA), (Softslip 12-well and Softwell 6-well) that were coated with 10 μg/mL fibronectin (Cultrex® #3420–001-01) in 1 × PBS for 1 h. All cells were allowed to sit on substrates for at least 12 h before BMP4 treatment.

Techniques: Immunofluorescence, Real-time Polymerase Chain Reaction, Control