Additional file 1: of Establishment of an erythroid progenitor cell line capable of enucleation achieved with an inducible c-Myc vector

Figure S1. Detailed DNA plasmid map of lentivirus transfer vector TRE3G-cMyc. Figure S2. Doubling time of c-Myc knock-out HO15.19 cells modified with the TRE3G-cMyc transfer vector compared with wild-typ type cells. Figure S3. Fluorescence activating cell sorting gates used to isolate erythroid enriched populations from fresh lineage depleted bone marrow. Figure S4. Colony forming cell (CFC) results for isolated Lin−c-Kit+CD71(low/−) mouse bone marrow cells. Figure S5. Comparing normal BFU-E and ‘BFU-E like’ colonies formed from normal and genetically modified hematopoietic cells. Figure S6. Cell surface protein profile of IPE cells at isolation and after culture. Figure S7. Cell surface protein profile of Lin−c-Kit+CD71(low/−) cells modified with the TRE3G-cMyc and cultured for three weeks. Supplementary Methods. Figure S8. Tetramethylbenzidine (TMB) staining of IPE cells after 48 h of culture with 0 ng/ml dox. Figure S9. Relative size of cells comparing IPE cells, RBCs derived from IPE cells (IPE - > RBC), and fresh RBCs. Figure S10. Example of gating strategy based on FSC and SSC and then on 7-AAD. Figure S11. Flowchart for IPE cell establishment, isolation, and optimised differentiation protocol. (DOCX 3550 kb)