Ectopic Expression of Ceacam1-L in G-2 Cells

For the ectopic expression of Ceacam1-L in G-2 cells, Ceacam1 cDNA was extracted from the pcDNA-mCC1-4 via EcoRI-digestion and subsequently cloned into lentiviral vector LeGO-iG2 (using EcoRI restriction site), kindly provided by Dr. C. Stocking [81 (link)]. Sequences were confirmed by DNA sequencing (Seqlab, Germany). Production of lentiviral vectors and transduction of G-2 cells was performed as previously described [12 (link)]. Commercially available shRNA constructs against mouse Ceacam1 mRNA using the pGIPZ vectorsystem were purchased at Open Biosystems (# RMM4532). The production of the lentiviral particles was performed according to the manufacturer instructions. GFP positivity was used to enrich cells that received the vector via fluorescence-activated cell-sorting.