Figure 2.

iC9-Expressing Cells Are Selectively Eradicated by AP1903

(A) Marking of H1-iC9 cells with GFP. Structure of the lentiviral vector containing the GFP gene is shown. H1-iC9 cells were transduced with the GFP vector at a multiplicity of infection of two, followed by cell sorting 48 h after transduction. GFP⁺ cells were pooled and expanded.

(B and C) Flow cytometry analysis of H1 and H1-iC9(GFP) cells either alone or in mixtures with and without AP1903 treatment as indicated. (B) FACS plots present the percentage of GFP⁺ cells in each group. (C) Bar graph shows mean (±s.d.) of pooled data from three independent experiments.

(D) Expression of iC9 transgene mRNA in the indicated cells. Results of three independent experiments are shown as the mean ± s.d.

(E) Experimental scheme for the teratoma formation assay. H1 and H1-iC9(GFP) cells were mixed at an equal dose and either mock treated or treated with AP1903 overnight at a concentration of 10 nM, followed by subcutaneous injection of 2 × 10⁶ cells into NSG mice with four mice per group. Teratomas were removed six weeks after injection.

(F) Representative immunofluorescence staining of GFP in teratoma sections derived from indicated cells. Scale bar = 50 μm.