Ab96792

Antibodies were purchased as follows: Primary antibodies: rabbit-anti-UCH-L1 (AB1761,MILLIPORE), rabbit-anti-synaptopodin (S9442, Sigma-Aldrich), mouse-anti-α-actinin4 (MAB1682, MILLIPORE), mouse-anti-nestin (MAB2763, R&D), rabbit-anti-NeuN (ab104225, Abcam), mouse-anti-S100 (ab14849, Abcam), rabbit-anti-NSE (ab53025, Abcam), mouse-anti-nephrin (sc-166574, Santa Cruz Biotechnology), mouse-anti-WT-1 (ab96792, Abcam). Secondary antibodies: peroxidase-conjugated goat-anti-mouse IgG, peroxidase-conjugated goat-anti-rabbit IgG, fluorescein (FITC)-conjugated goat-anti-rabbit/mouse IgG, cy3-conjugated goat-anti-mouse/rabbit IgG all from Protein Tech Group Inc., Chicago, USA, HRP-goat-anti-mouse IgG, HRP-goat-anti-rabbit IgG from Long Island Biotech Co., LTD, Shanghai, China.

For Western-blots, total proteins were extracted from tissues or cultured cells using RIPA buffer containing protease inhibitors cOmplete, ULTRA, Mini, EDTA-free, EASYpack (Roche, Basel, Switzerland), while the membrane proteins were extracted from tissues by Mem-PER Eukaryotic Membrane Protein Extraction Reagent Kit (Thermo Scientific, Rockford, USA). Protein concentrations were determined by the BCA method. Equal amount of proteins (100ug) were separated with 7.5 %/12.5 % sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to PVDF membrane. Membrane was blocked using 5 % skimmed milk and incubated with respective antibodies. Primary polyclonal antibodies were purchased from Santa Cruz Biotechnology (CA, USA) and Abcam (CA, USA) including WT1 (ab96792), p-STAT3 (ab30647), STAT3 (ab68153), p-MAPK (ab63378), MAPK (ab197348), GAPDH (sc-48166), β-actin (sc-47778), Caspase 3 (ab2302), MDR1 (sc-13131). The secondary antibodies were anti-rabbit or anti-mouse HRP-linked were purchased from Santa Cruz Biotechnology (CA, USA). The blots were developed using ECL reagent (Millpore, MASS, USA). Equal amount of protein loading in each lane was confirmed using GAPDH antibody. All experiments were repeated at least three times.