Nuclear localization of intracellular domain of LDL receptor-related protein 1B induced expresion data

We have generated a MCF-7 Tet-On model system for inducible nuclear expression of intracellular domain of LRP1b using the pTRE-Tight vector system (Clontech Takara, Ohtsu, Japan). Expression data of MCF-7 without and with induction of Doxycycline, sample 1 and sample 2, respectively. We have generated a MCF-7 Tet-On model system for inducible expression of intracellular domain of LRP1b using the pTRE-Tight vector system (Clontech Takara, Ohtsu, Japan). Human cDNA, which encoded a intracellular lesion of LRP1B, was amplified by PCR using sense primer 5’-ACCATGGGTAAAAGAAAAAGAAGGACAAAAAC-3’ (underline indicates the introduced start codon) and antisense primer 5’-AGGGATCCTCACTGATTATGCCACTGTCTCTCTTATAC-3’(double underline and underline indicates the introduced BamH1 site and internal stop codon, respectively), cloned to pTarget-T vector (Promega), and subsequently verified by sequencing. Next, Mlu1 and BamH1 fragment, which contained the coding region of intracellular domain of LRP1B was subcloned to Mlu1-BamH1 site of pTRE3G-ZsGreen1 vector (Clontech Takara). After verified by sequencing of the inserted lesion, the vector was designated pTRE3G-ZsGreen1-cLRP1B vector. A stable Tet-On 3G MCF-7 cell line, which was a host for Tet-inducible gene expression systems, was established using a pCMV-Tet3G vector and Xfect transfection reagent (Clontech Takara), subsequently transfected with pTRE3G-ZsGreen1-cLRP1B vector according to the manufacture’s protocol. Finally, a doxycycline (Dox) controlled expression of intracellular domain of LRP1B was established in MCF-7 cells. We established six MCF cell clones, in which intracellular domain of LRP1B was tightly regulated by doxycycline (Dox) at 100ng/ml.