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 .