Figure 4
Figure 4. Relationships among VWF, GPIIb, GPIIIa, β-actin, GAPDH, β2-microglobulin, CD34 and CD71 protein level, and ploidy. The levels of (A) VWF, GPIIb, GPIIIa, (B) β-actin, GAPDH, β2-microglobulin, and (C) CD34 and CD71 in each ploidy class were measured by flow cytometry. (i) Geometric mean fluorescence intensity (MFI) was calculated for each ploidy class as follows. Each ploidy class was first gated (from 2N to 16N) on the propidium iodide (PI) labeling. MFI in FL1 was measured for each specific antibody and its isotype-matched control (IgG). The corrected MFI for each ploidy was calculated by subtraction of the isotype MFI to the MFI obtained with the specific antibody. (ii) Examples of MFI measurement. Cells were stained with specific antibody and PI and gated on the ploidy class (middle panels). MFI designed as M1 for IgG and as M2 for (A) GPIIb, (B) β-actin, and (C) CD71, respectively, was obtained for each ploidy level separately by measuring the fluorescence intensity in FL1 (lower panels). Corrected MFI is equal to the subtraction of M1 from M2. Filled areas represent background fluorescence (M1), whereas open areas represent fluorescence obtained with the specific antibody (M2).

Relationships among VWF, GPIIb, GPIIIa, β-actin, GAPDH, β2-microglobulin, CD34 and CD71 protein level, and ploidy. The levels of (A) VWF, GPIIb, GPIIIa, (B) β-actin, GAPDH, β2-microglobulin, and (C) CD34 and CD71 in each ploidy class were measured by flow cytometry. (i) Geometric mean fluorescence intensity (MFI) was calculated for each ploidy class as follows. Each ploidy class was first gated (from 2N to 16N) on the propidium iodide (PI) labeling. MFI in FL1 was measured for each specific antibody and its isotype-matched control (IgG). The corrected MFI for each ploidy was calculated by subtraction of the isotype MFI to the MFI obtained with the specific antibody. (ii) Examples of MFI measurement. Cells were stained with specific antibody and PI and gated on the ploidy class (middle panels). MFI designed as M1 for IgG and as M2 for (A) GPIIb, (B) β-actin, and (C) CD71, respectively, was obtained for each ploidy level separately by measuring the fluorescence intensity in FL1 (lower panels). Corrected MFI is equal to the subtraction of M1 from M2. Filled areas represent background fluorescence (M1), whereas open areas represent fluorescence obtained with the specific antibody (M2).

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