Rabbit anti jagged1

SB203580, SP600125, PD98059 and PDTC were purchased from Calbiochem and used at concentration of 10 μM. Rabbit anti-Notch4 (sc-5594, 1:200 dilution) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA); Rabbit anti-Jagged1 (2620, 1:000 dilution) from Cell Signaling Technology; Rabbit anti-GAPDH (SAB2701826) from Sigma. PGN were purchased from Sigma.

Cells were harvest in lysis buffer on ice. Sample proteins were loaded on SDS-PAGE gel and blotted onto PVDF membranes. After blocking in 5% (w/v) non-fat milk for 1 h, proteins were probed with primary antibodies at 4 °C overnight. Primary antibodies were diluted in 5% bovine serum albumin blocking buffer as follows: rabbit anti-Jagged1 (Cell Signalling Technology, USA; 1:500), rabbit anti-Notch1 (Cell Signalling Technology, USA; 1:1000), mouse anti-MBP (Biolegend, USA; 1:1000). Membranes were then incubated in horseradish peroxidase-conjugated secondary antibody for 1 h, the immunoreactive proteins can be detected by chemiluminescence ECL agents (Millipore, USA).