Figure 2
Figure 2. Epo-specific PODXL induction in developmentally staged marrow erythroblasts. (A) In MACS-purified Kit+CD71high erythroblasts, time courses of EPO and SCF induction of PODXL expression (after cytokine withdrawal) were assessed by quantitative RT-PCR. (B) In SP34-EX expansion cultures, cell-surface PODXL expression among Kit+CD71high, Kit−CD71high, and Kit−CD71highTer119+ erythroblasts was analyzed by flow cytometry. Frequencies of PODXL+ cells are indicated for bisected PODXLlow and PODXLhigh subpopulations. (C) For the above stages of maturing erythroblasts, cytospin morphologies for MACS and/or FACS-purified populations also are shown. (Images were seen with a Zeiss (Oberkochen, Germany) model Axioskop 2 microscope with a 100 ×/1.3 oil lens, Immersol 518 (Zeiss) imaging solution, and Volu-sol (Volu-Sol Inc.) dip stain solution. Images were photographed with a Spotflex (Diagnostic Instruments Inc., Sterling Heights, MI) model 15.2 camera and processed with Spot advance (Diagnostic Instruments Inc.) version 4.6 and Photoshop (Adobe, San Jose, CA) version CS2 software.

Epo-specific PODXL induction in developmentally staged marrow erythroblasts. (A) In MACS-purified Kit+CD71high erythroblasts, time courses of EPO and SCF induction of PODXL expression (after cytokine withdrawal) were assessed by quantitative RT-PCR. (B) In SP34-EX expansion cultures, cell-surface PODXL expression among Kit+CD71high, KitCD71high, and KitCD71highTer119+ erythroblasts was analyzed by flow cytometry. Frequencies of PODXL+ cells are indicated for bisected PODXLlow and PODXLhigh subpopulations. (C) For the above stages of maturing erythroblasts, cytospin morphologies for MACS and/or FACS-purified populations also are shown. (Images were seen with a Zeiss (Oberkochen, Germany) model Axioskop 2 microscope with a 100 ×/1.3 oil lens, Immersol 518 (Zeiss) imaging solution, and Volu-sol (Volu-Sol Inc.) dip stain solution. Images were photographed with a Spotflex (Diagnostic Instruments Inc., Sterling Heights, MI) model 15.2 camera and processed with Spot advance (Diagnostic Instruments Inc.) version 4.6 and Photoshop (Adobe, San Jose, CA) version CS2 software.

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