Western blot was performed as common methods. The protein of HUVEC cells was
isolated in radioimmunoprecipitation (RIPA) buffer supplemented with PMSF
(Beyotime). The protein were separated by using 10% SDS-PAGE gels and
immediately moved onto polyvinylidenedifluoride membranes (PVDF; Millipore,
Boston, MA, USA) via a wet transfer method. Subsequently, these membranes were
incubated in specific antibodies at 4°C overnight. On the next day, protein band
visualization was implemented by exposing to enhanced chemiluminescent (ECL)
reagent after incubated with secondary antibodies. GRP 78 antibodies (Abcam, Cat
No: ab21685), ATF 6 antibodies (Abcam, Cat No: ab203119) and primary antibody:
CD63, TSG-101, LAMC1, β-catenin, c-myc, cyclin D1 and GAPDH (Cell Signaling
Technology, Danvers, MA, USA).
isolated in radioimmunoprecipitation (RIPA) buffer supplemented with PMSF
(Beyotime). The protein were separated by using 10% SDS-PAGE gels and
immediately moved onto polyvinylidenedifluoride membranes (PVDF; Millipore,
Boston, MA, USA) via a wet transfer method. Subsequently, these membranes were
incubated in specific antibodies at 4°C overnight. On the next day, protein band
visualization was implemented by exposing to enhanced chemiluminescent (ECL)
reagent after incubated with secondary antibodies. GRP 78 antibodies (Abcam, Cat
No: ab21685), ATF 6 antibodies (Abcam, Cat No: ab203119) and primary antibody:
CD63, TSG-101, LAMC1, β-catenin, c-myc, cyclin D1 and GAPDH (Cell Signaling
Technology, Danvers, MA, USA).