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Fig. 4 | Journal of Biomedical Science

Fig. 4

From: WSB1: from homeostasis to hypoxia

Fig. 4

Preparation of Tet-inducible-WSB1 Cell Lines. Full length wsb1 isoform 1 was prepared by RT-PCR and cloned into a shuttle vector. Clones were sequenced and both SNP variants C and T were identified and each was cloned in frame, into the Tet-On 3G system vector, pTRE3G (Clontech). The wsb1 insert in pTRE3G was sequenced prior to use for transfection. wsb1-pTRE3G was transfected into either HeLa or HEK293 containing a stably transfected TET protein expressing construct (Clontech). Puromycin resistant clones were selected and evaluated for their inducibility with 1 mg/ml doxycycline. Figure 4 demonstrates the inducibility of wsb1 in the presence or absence of doxycycline. Example cell lines include either WSB1 “C” variant HEK293 (2 clones), or “T” variant (one cell line) or HeLa “T” variant

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