Anti goat and mouse

Cell lysates were prepared in 8 M urea (Sigma-Aldrich), protein concentration was determined by Biorad Protein Assay (Bio-Rad, Munchen, Germany). Equal amounts of proteins (40 μg) were subjected to SDS–polyacrylamide gel electrophoresis and were transferred onto a nitrocellulose membrane saturated with 5% bovine serum albumin (BSA) in Tris-buffered saline with 0.1% Tween 20. Membranes were incubated with primary antibody for 18 h at 4°C and subsequently with horseradish peroxidase-conjugated secondary antibody for 1 h at room temperature. Membranes were washed with Tris-buffered saline in 0.1% Tween 20 and developed using the chemiluminescence system Chemidoc MP Imaging System (Bio-Rad). Antibody anti-p21, -p53, -SHMT2, -β-actin and the secondary antibodies anti-goat and -mouse were from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Cell lysates were prepared in 8 M urea (Sigma-Aldrich) and protein concentration was determined by Biorad Protein Assay (Bio-Rad, Munchen, Germany). Lysates were subjected to SDS–polyacrylamide gel electrophoresis and were transferred onto a nitrocellulose membrane then saturated with 5% non fat dry milk in Tris-buffered saline with 0.1% Tween-20 for 1 h at room temperature. Membranes were incubated with primary antibody overnight at 4°C and subsequently with horseradish peroxidase-conjugated secondary antibody for 1 h at room temperature. Membranes were washed with Tris-buffered saline with 0.1% Tween-20 and ECL reagent (Millipore) was used for detection by chemiluminescence system Chemidoc MP Imaging System (Bio-Rad). Antibody anti-SHMT1 were from Cell Signaling Technology (Danvers, MA, USA), the secondary antibody anti-rabbit were from Abbiotec (San Diego, CA, USA), antibody anti-SHMT2, -β-actin and the secondary antibodies anti-goat and -mouse were from Santa Cruz Biotechnology (Santa Cruz, CA, USA).