Hrp conjugate immunoglobulin

Western blot analysis was performed as previously described (Chen Y. et al., 2020 (link)). Briefly, cell lysates were separated on 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gels and then transferred onto nitrocellulose membranes. Sources and dilution of antibodies were shown in Table 2. HRP conjugate immunoglobulin was used as a secondary antibody (Jackson ImmunoResearch Laboratories, West Grove, PA, United States). West Pico chemiluminescent (Pierce) was used as the substrate to visualize protein bands, which were quantified using densitometric image analysis software (Image Master VDS; Pharmacia Biotech). Normalization was made against GAPDH.

Twenty micrograms of cell lysates were separated on 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gels and then transferred onto nitrocellulose membranes. Specific monoclonal anti-Notch1 (CST, SN: 4380, delution: 1:2000), monoclonal anti-Hes1 (CST, SN: 11988, delution: 1:2000), monoclonal anti-Vimentin (CST, SN: 5741, delution: 1:2000), monoclonal anti-E-cadherin (CST, SN: 3195, delution: 1:2000) and monoclonal anti-β-actin (CST, SN: 8457, delution: 1:4000) were used. HRP conjugate immunoglobulin was used as a secondary antibody (Jackson ImmunoResearch Laboratories, West Grove, PA, USA). West Pico chemiluminescent (Pierce) was used as the substrate to visualize protein bands, which were quantified using densitometric image analysis software (Image Master VDS; Pharmacia Biotech). Normalization was made against β-actin expression.