Development of GFP-IBD Reporter Cell Lines

The MCF7 Tet-On Advanced cell line was obtained from Clontech. The generation and characterization of the MCF7^GFP-IBD cell line has been described (24 (link)) and was used with further authentication by IDEXX BioResearch within the last 6 months. Panc 02^GFP-IBD, U-87 MG^GFP-IBD, and hTERT-HME1^GFP-IBD cell lines were developed similarly from parent cell lines purchased from American Type Culture Collection (ATCC). Briefly, GFP-IBD cloned into the pLVX-Tight-Puro vector was transfected along with pLVX-Tet-On Advanced vector (Clontech) into each cell line. Following G418 and puromycin selection, cells were induced with 1 μg/mL doxycycline and sorted to establish the IRIF reporter cell lines. The Panc 02^GFP-IBD and U-87 MG^GFP-IBD cell lines were maintained in RPMI (Invitrogen), supplemented with 10% Tet system approved FBS (Clontech). The hTERT-HME1^GFP-IBD cell line was maintained in MEBM media supplemented with MEGM SingleQuot (Lonza). For studies requiring glucose and glutamine limitation, media was prepared using DMEM base, D-glucose (Sigma), and L-glutamine solutions (Gemini Bioproducts) at appropriate concentrations with 10% FBS.