Tropix cspd

Protein extracts were isolated using TRIzol Reagent (Invitrogen) following manufacturer´s indications, clarified (12,000 × g, 15 min, 4°C), denatured and subjected to SDS-PAGE and Western-blot. Antibodies used were rabbit polyclonal anti-SET (Abcam) and mouse monoclonal anti-βactin (Sigma). Proteins were detected with the appropriate secondary antibodies conjugated to alkaline phospatase (Sigma) by chemiluminescence using Tropix CSPD and Tropix Nitro Block II (Applied Biosystems).

Protein extracts were isolated using TRIzol Reagent (Invitrogen) following the manufacturer’s indications, denatured and separated on a 10% SDS-PAGE and western blot. Rabbit polyclonal anti-FoxP3 (Abcam) and mouse monoclonal anti-β-actin (Sigma) antibodies were used. Proteins were detected with the appropriate secondary antibodies conjugated to alkaline phosphatase (Sigma) by chemiluminescence using Tropix CSPD and Tropix Nitro Block II (Applied Biosystems).

Protein extracts were isolated using TRIzol Reagent (Invitrogen, Carlsbad, CA, USA) following the manufacturer’s indications; these were then clarified (12,000× g, 15 min, 4 °C), denatured, and subjected to SDS-PAGE and western blotting. The antibodies used were mouse monoclonal anti-PP2A (Upstate Inc., Lake Placid, NY, USA), rabbit monoclonal anti-p-PP2A-C Y307 (Epitomics, Burlingame, CA, USA), rabbit polyclonal anti-AKT, rabbit polyclonal anti-ERK (Cell Signaling Technology Inc., Beverly, MA, USA), rabbit polyclonal anti-SET (Abcam, Cambridge, MA, USA), rabbit polyclonal anti-pAKT^Thr308, rabbit polyclonal anti-pERK1/2^{Thr202/Tyr204} (Santa Cruz Biotechnology, Santa Cruz, CA, USA), rabbit polyclonal anti-CIP2A, and mouse monoclonal anti-β actin (Sigma, St. Louis, MO, USA). Proteins were detected with the appropriate secondary antibodies conjugated to alkaline phospatase (Sigma, St. Louis, MO, USA) by chemiluminescence using Tropix CSPD and Tropix Nitro Block II (Applied Biosystems, Foster City, CA, USA).