Anti sox11

Proteins were extracted using RIPA buffer (Sigma), supplemented with protease (1:1000, Sigma) inhibitor cocktails, after 30 min incubation on ice. Protein concentrations were measured using BCA assay, with BSA for the standard curve, and 50 μg of protein were resolved on 12% NuPAGE gels (Invitrogen) and transferred using a semi-dry transfer system onto PVDF membranes (Millipore). Non-specific binding sites were blocked with 10% milk TBS-T solution for 1 h at room temperature, then probed overnight at 4 °C with the respective primary antibodies: anti-SAMHD1 (Abcam), anti-SOX11 (Sigma), or anti-Cyclin D1 1:1000 in 5% milk or 5% BSA in TBS-T. Membranes were then washed in TBS-T and probed with secondary antibodies (HRP-conjugated anti-rabbit or anti-mouse; GE Healthcare). Blots were developed using Supersignal West Pico (Pierce) and visualized using LiCor machine. For re-probing of membranes with anti-actin (Sigma) or anti-GAPDH (Cell Signaling) (1:5000 in 5% milk in TBS-T, Sigma), HRP was blocked using the SG substrate kit (Vector Labs). Analysis was done using Fiji-ImageJ software.

Anti-β-catenin and anti-p27^kip1 antibodies were purchased from BD Biosciences (San Jose, CA, USA). Anti-Sox4, anti-Sox6, anti-Sox7, anti-Sox9, anti-Sox11, and β-actin antibodies were obtained from Sigma-Aldrich Chemicals (St. Louis, MO, USA). Anti-Snail and anti-Slug antibodies were from Cell Signaling (Danvers, MA, USA). Anti-p21^waf1, anti-cyclin D1, and anti-CD44s antibodies were purchased from Dako (Copenhagen, Denmark). Anti-Sox2 and anti-cyclin A antibodies were from Abcam (Cambridge, MA, USA) and Novocastra (Newcastle, UK), respectively. Anti-HA and anti-E-cadherin antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA) and Takara (Shiga, Japan) respectively. Anti-CD133 antibody was from Miltenyi Biotechnology (Bergisch Gladbach, Germany).
STK2, which is a serum-free culture medium for mesenchymal stem cells, [22 (link)] was obtained from DS Pharma Biomedical (Osaka, Japan).