Ubiquitination assay was carried out in HEK293T cells. In HEK293T cells, HA-Ub and Flag-YAP1 were co-transfected with empty vector, Myc-PDLIM7 or Myc-TPM2 plasmids. Cells were treated with 10 mM MG132 for 6 h before harvest. Ubiquitinated YAP1 proteins were enriched by anti-Flag magnetic beads. After eluted with the Flag peptide, ubiquitinated YAP1 was detected by anti-HA antibody.
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