Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Phospho-YAP1 (S127) antibody (ET1611-69) at 1/5,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1611-69) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Western blot analysis of Phospho-YAP1 (S127) on different lysates with Rabbit anti-Phospho-YAP1 (S127) antibody (ET1611-69) at 1/5,000 dilution. Lane 1: HeLa whole cell lysate Lane 2: HeLa treated with 100nM Calyculin A for 30 minutes whole cell lysate Lane 3: HeLa treated with 100nM Calyculin A for 30 minutes then treated with λpp for 1 hour whole cell lysate Lane 4: NIH/3T3 whole cell lysate Lane 5: NIH/3T3 treated with 100nM Calyculin A for 30 minutes whole cell lysate Lane 6: NIH/3T3 treated with 100nM Calyculin A for 30 minutes then treated with λpp for 1 hour whole cell lysate Lane 7: C6 whole cell lysate Lane 8: C6 treated with 100nM Calyculin A for 30 minutes whole cell lysate Lane 9: C6 treated with 100nM Calyculin A for 30 minutes then treated with λpp for 1 hour whole cell lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 54 kDa Observed band size: 70 kDa Exposure time: 1 minute 6 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1611-69) at 1/5,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.