Immunostaining was performed by using mouse monoclonal antibodies to OATP1B3 (Novus-74482, Novus Biologicals), OATP1B1 (Abcam42, andaMRP2ab3373P2 (Abcam3, Abcam).
PR was detected by using mouse monoclonal anti-PR (1 50; hPRa7; Neomarkers, Fremont, CA, USA).
Immunohistochemical procedures were performed on all tissue by using mouse monoclonal antibody to the nucleoprotein of influenza A H5N1 (B.V. European Veterinary Laboratory, Woerden, the Netherlands).
Inhibition of the proliferation of PBMC to AQP4 p61 80 and rhAQP4 was studied by using mouse monoclonal anti HLA-DR (clone G46-6; BD Biosciences), anti HLA-DR (clone HG46-6 anti HLA-DQridge, Manti HLA-DQA-DP (clone B7/21; Abcam), and isotype control (clone G155-178; BD Biosciences).
Sections of all patients were stained with T cell (CD2, CD3, and CD45) and B cell (CD20 and CD79a) markers by using mouse monoclonal antibodies, and disease specificity was confirmed in case that positive staining was observed only with one or more T cell-specific markers without any positive B cell-specific marker staining.
SOX11 expression was detected by using a rabbit monoclonal antibody (Epitomics, Burlingame, CA, USA, clone EPR8192); caspase-3 (R&D Systems, Minneapolis, MN, USA) and cleaved caspase-3 (Cell Signaling Technology, Danvers, MA, USA) were detected by using mouse monoclonal and rabbit polyclonal antibodies.
C. perfringens alpha toxin (1 µg/ml) purchased from Sigma Aldrich, or culture supernatants, were analyzed by ELISA using mouse monoclonal anti-CPA antibody (kindly provided by Dr. Paul Hauer), as previously described [43].
Ubiquitin was detected by immunoblotting using mouse monoclonal antibody against ubiquitin (1 : 1000; Santa Cruz, Heidelberg, Germany).
HA-tagged Wp85 and Δp85 proteins in the transfectants were detected by immunoblotting using mouse monoclonal anti-HA antibodies (Santa Cruz Biotechnology, Inc).
Remaining immunoprecipitates were separated on 10 20% (w/v) Tris/HCl pre-casted gels and analyzed by immunoblotting using mouse monoclonal anti-RhoA antibody and HRP-conjugated mouse secondary antibody.
ER status was determined by immunohistochemistry using mouse monoclonal anti-human ER antibody (Dako, Carpinteria, CA, clone 1D5) on Chinese tumors or using ER-ICA (estrogen receptor immunocytochemical assay) assay on Italian tumors (Abbott, Abbott Park, IL) according to the manufacturer's recommendations.
