Anti active β catenin 8e7

Cells were fixed with 4% paraformaldehyde and blocked with blocking buffer (1 × PBS/5% goat serum/0.3% Triton X-100). Cells were labeled with anti-active-β-catenin (8E7, Merck Millipore, Billerica, MA, United States) at 4°C overnight. After washing the cells, cells were incubated with secondary antibodies (Alexa Fluor 555 anti-mouse IgG, Thermo Fisher Scientific) at room temperature for 1 hr. After washing the cells, active-β-catenin was observed under the fluorescence microscope (EVOS Cell Imaging System, Thermo Fisher Scientific).

The following antibodies (sources indicated between brackets) were used: anti‐DDR1 C‐20, anti‐β‐catenin E‐5 (Santa Cruz Biotechnology); anti‐ERK1/2, anti‐ERK1/2 pT202/Y204, anti‐AKT, anti‐AKT pS473 anti‐pY792 DDR1, anti‐non‐phospho (active) β‐catenin (Ser33/37/Thr41), anti‐BCR N‐20 and anti‐pTyr clone pY1000 Sepharose bead conjugated (PTM Scan, Cell Signaling Technology); anti‐β‐actin (Sigma‐Aldrich); anti‐active‐β‐catenin 8E7 (Merck Millipore); anti‐pY177 BCR (Abgent); anti‐tubulin (gift from N. Morin, CRBM, Montpellier, France); anti‐pTyr 4G10 (gift from P. Mangeat, CRBM, Montpellier, France); anti‐rabbit IgG‐HRP and anti‐mouse IgG‐HRP (GE Healthcare); goat anti‐mouse Alexa Fluor 594 (Thermo Fisher Scientific). Nilotinib was a generous gift from Novartis, and collagen I rat tail was from Invitrogen and IWR‐1‐endo from SelleckChem.