Ab92299

All TMA and conventional sections were deparaffinized at 60 °C for 20 min and rehydrated in graded ethanols. Antigen retrieval was performed by immersing the tissues in citrate buffer (pH = 6.0) for 20 min in a water bath. Endogenous peroxidase and non-specific staining were blocked with 3% H₂O₂ for 20 min at room temperature. The TMA sections were incubated for 1 hour at room temperature with the following primary antibodies: PRMT1 (ab92299, Abcam, dilution: 1/200), ZEB1 (HPA027524, Sigma Aldrich, dilution: 1/500), RUNX1 (sc-365,644, Santa Cruz, dilution: 1/50), TWIST1 (ABD29 Merck Millipore, dilution: 1/100), E-cadherin (spm471, Santa Cruz, dilution: 1/100), N-cadherin (6D11, DAKO, dilution: 1/100), ß-catenin (ß-catenin-1, Dako, dilution: 1/100). Whole-mount sections were incubated with the PRMT1 (ab92299, Abcam, dilution: 1/200) and ZEB1 (HPA027524, Sigma Aldrich, dilution: 1/500). All slides were then incubated with Envision ⁱⁿ FLEX/HRP (K4063, Dako, Denmark) for 30 min. Staining patterns were visualized by exposure to 3,3′-diaminobenzidine (DAB) and counterstained with Mayer’s hematoxylin. Finally, the slides were dehydrated in ethanol, cleared in xylene and mounted for examination. Human kidney tissue was used as a positive control of staining and replacement of the primary antibody with Tris-buffered saline was used as a negative control.