American Society of Hematology

Fig. 2.

$Fig. 2. FACS profiles of transduced +/+ and IL-3−/− BM cells. / (A) Dot plot showing gates used to isolate the Sca-1+lin− subset of GFP+ cells after exposure of +/+ and IL-3−/− BM cells to BCR-ABL-GFP. (B) Dot plot showing gates used to isolate the GFP+(BCR-ABL+, lower right), YFP+(IL-3+, upper left), and GFP+ and YFP+ (BCR-ABL+ andIL-3+, upper right) cells within the lin−subset after simultaneous exposure of +/+ and IL-3−/− BM cells to BCR-ABL-GFP and IL-3-YFP virus. (C) RT-PCR analysis of RNA isolated from FACS purified GFP+, YFP+, and GFP+ and YFP+ fractions of cells shown in Figure 1B. Lanes 1 and 5 show results for MIG-transduced GFP+ IL-3−/− and +/+ cells; lanes 2 and 6 for cells cotransduced with BCR-ABL and IL-3 and purified for YFP (IL-3) expression only; lanes 3 and 7 for cells cotransduced with BCR-ABL and IL-3 and purified for GFP (BCR-ABL) expression only; and lanes 4 and 8 for cells cotransduced with BCR-ABL and IL-3 and purified for dual GFP and YFP (BCR-ABL and IL-3) expression. Lanes 9 and 10 were water controls (negative controls) for the RT and PCR reactions.$

FACS profiles of transduced +/+ and IL-3^−/− BM cells.

(A) Dot plot showing gates used to isolate the Sca-1⁺lin⁻ subset of GFP⁺ cells after exposure of +/+ and IL-3^−/− BM cells to BCR-ABL-GFP. (B) Dot plot showing gates used to isolate the GFP⁺(BCR-ABL⁺, lower right), YFP⁺(IL-3⁺, upper left), and GFP⁺ and YFP⁺ (BCR-ABL⁺andIL-3⁺, upper right) cells within the lin⁻subset after simultaneous exposure of +/+ and IL-3^−/− BM cells to BCR-ABL-GFP and IL-3-YFP virus. (C) RT-PCR analysis of RNA isolated from FACS purified GFP⁺, YFP⁺, and GFP⁺ and YFP⁺ fractions of cells shown in Figure 1B. Lanes 1 and 5 show results for MIG-transduced GFP⁺ IL-3^−/− and +/+ cells; lanes 2 and 6 for cells cotransduced with BCR-ABL and IL-3 and purified for YFP (IL-3) expression only; lanes 3 and 7 for cells cotransduced with BCR-ABL and IL-3 and purified for GFP (BCR-ABL) expression only; and lanes 4 and 8 for cells cotransduced with BCR-ABL and IL-3 and purified for dual GFP and YFP (BCR-ABL and IL-3) expression. Lanes 9 and 10 were water controls (negative controls) for the RT and PCR reactions.

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