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EM_21724_sm_SuppFig1.TIF78KSupplemental Fig. 1 The distribution of singlets and doublets of red blood cells (RBCs). A cell population was plotted by the area signal of forward scatter vs. the peak signal of forward scatter. Singlets were distributed through the lower gated region and doublets were distributed through the upper gated region. This data were derived from a unirradiated mouse.
EM_21724_sm_SuppFig2a.TIF151KSupplemental Fig. 2 Spheroidizing of RBCs. After washing, the stained blood solution was divided in half. One solution was analyzed directly and the other solution was added to about a 25-times volume of 0.003% sodium dodecyl sulfate (SDS) in phosphate-buffered saline (PBS), which was subsequently added to about a 25-times volume of 0.001% SDS and 0.2% glutaraldehyde in PBS. (A)(B) The data without the spheroidizing of the RBCs and after the spheroidizing of the RBCs are shown in the upper left and upper right of the TER-119 parameter histogram, respectively. The lower left dot plot cytogram was plotted by fluorescein isothiocyanate-conjugated anti-mouse CD24 antibody (anti-CD24-FITC) versus phycoerythrin anti-mouse CD71 antibody (anti-CD71-PE) fluorescence without the spheroidizing of RBCs and the lower right dot plot cytogram was plotted after spheroidizing of RBCs. The values located in the upper right of each quadrant (ie, B1, B2, B3, and B4) indicate the number of events detected. (A) shows the data from a mouse irradiated with 2 Gy of X-rays 2 weeks after dosing and (B) shows the data from a mouse treated with 142 mg/kg N-Ethyl-N-nitrosourea (Sigma-Aldrich Co., St. Louis, MO, USA) 2 weeks after dosing.
EM_21724_sm_SuppFig2b.TIF154KSupplemental Fig. 2 Spheroidizing of RBCs. After washing, the stained blood solution was divided in half. One solution was analyzed directly and the other solution was added to about a 25-times volume of 0.003% sodium dodecyl sulfate (SDS) in phosphate-buffered saline (PBS), which was subsequently added to about a 25-times volume of 0.001% SDS and 0.2% glutaraldehyde in PBS. (A)(B) The data without the spheroidizing of the RBCs and after the spheroidizing of the RBCs are shown in the upper left and upper right of the TER-119 parameter histogram, respectively. The lower left dot plot cytogram was plotted by fluorescein isothiocyanate-conjugated anti-mouse CD24 antibody (anti-CD24-FITC) versus phycoerythrin anti-mouse CD71 antibody (anti-CD71-PE) fluorescence without the spheroidizing of RBCs and the lower right dot plot cytogram was plotted after spheroidizing of RBCs. The values located in the upper right of each quadrant (ie, B1, B2, B3, and B4) indicate the number of events detected. (A) shows the data from a mouse irradiated with 2 Gy of X-rays 2 weeks after dosing and (B) shows the data from a mouse treated with 142 mg/kg N-Ethyl-N-nitrosourea (Sigma-Aldrich Co., St. Louis, MO, USA) 2 weeks after dosing.
EM_21724_sm_SuppInfo.docx13KSupporting Information

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