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| Protein Electrophoresis and Western | Cells were lysed on ice for 20 min and supernatant was collected after centrifugation at 21,000g for 30 min at 4 °C. Cell lysate was later added 1× loading buffer (10% glycerol, 2% SDS, 0.05 M Tris-HCl pH 6.4, 0.01 M 2-mercaptoethanol, 0.6% bromophenol blue), and denatured at 90 °C for 10 mins. A 10 µl mixture was then added to SurePAGETM 4–12% Bis-Tris gels (GenScript, M00654). | Get A Quote |
TEA domain transcription factor 1 (TEAD1), a Hippo pathway transcription factor important in cellular homeostasis and development, is increasingly implicated in cancer biology. Here we reveal an unexpected role for TEAD1 in organizing nuclear condensates, independent of active transcription. Using high-resolution imaging, ChIP–seq, RNA sequencing and proximity-based proteomics, we demonstrate that in patient-derived renal cell carcinoma cells, TEAD1 forms micrometre-sized condensates by binding to heterochromatic pericentromeric regions using its DNA-binding domain. TEAD-specific MCAT motifs selectively enrich and cluster in the pericentromeric region to specifically seed TEAD1 condensates. TEAD1 condensate... More
