Constructs and cell culture
Detected gene variants were introduced into a pMT2-VWF expression vector
containing human wild-type (wt) VWF cDNA using site-directed mutagenesis
. HEK 293T (DSMZ, Germany) and HEK293 (ATCC, USA) cells were cultured
and transfected by lipofectamine (Life Technologies, USA), as described
previously , to evaluate the constitutive secretion and intracellular
localization of VWF, respectively .