L3MBTL1-KD leads to expansion of erythroid progenitors in long-term culture. (A) Sorted 4 × 105 GFP+ CD34+ were plated on MS5 stromal cell layer and cultured for 5 weeks. At week 5, the colonies were examined using an inverted optical microscope. The red arrows indicate the cobblestone area forming cells in the wt and shLUC figures. The red arrows indicate the overgrowth of progenitor cells (predominantly erythroid) in the sh1 and sh2 photographs. (B) The expression of GlyA on floating cells from 5-week LTC-IC cultures was evaluated by flow cytometry. (C) CAFC colony numbers were evaluated at week 5 of MS-5 stromal cell–based culture (n = 2). (D) Week 5 LTC-IC cells were plated on methylcellulose, and the secondary BFU-E colonies were scored after 10 days. The ratio of BFU-E colonies is shown, based on BFU-E numbers in the control cells.
