Expression of WT NE and the G185R mutant in HL-60 cells. (A) Schematic representation of mismatched PCR showing the polymorphism at S173 used to distinguish endogenous NE from untransduced cells (UNT) and ectopically expressed WT NE and the G185R mutant. (B) StuI restriction analysis of reverse transcription-PCR (RT-PCR) products demonstrating the presence of ectopically expressed WT NE or the G185R mutant (130 bp) and endogenously expressed NE (151 bp) in expanded clones. The negative image of an ethidium bromide-stained agarose gel is shown. Water (H₂O) indicates negative control; +CTRL, pCDNA3.1 plasmid containing NE as a positive control; VEC, cells transduced with empty MIEG3 vector. (C) Flow cytometric analysis of expanded clones demonstrating more than 96% EGFP-positive cells. M1 and M2 represent negative and positive gates for EGFP fluorescence, respectively.