Figure 1
Figure 1. Analysis of GPC sorting during enucleation of WT and 4.1R-null erythroblasts. Differential interference contrast (DIC) and immunofluorescent micrographs of wild-type (WT) and 4.1R-null enucleating erythroblasts, including nascent reticulocyte (R) and extruding nucleus (N), probed with fluorescein isothiocyanate–conjugated TER 119, specific for GPA (green), or Alexa Fluor 555–labeled rabbit anti–mouse GPC antibody (red). Nuclei were identified by Syto-17 staining (blue). The number of enucleating erythroblasts examined under each staining condition was 6 or more. The images were observed by the use of a Zeiss LSM META 510 Confocal microscope (Carl Zeiss Microimaging Inc) with an APOCHROMAT 63×/1.4 oil DIC objective and acquired by the use of Zeiss Laser Scanning Microscope LSM 510, Version 3.2 SP2 software with a Zeiss AxioCam HRm Rev. 2/3.3V camera. The images were processed with the use of Adobe Photoshop (Adobe Systems Inc).

Analysis of GPC sorting during enucleation of WT and 4.1R-null erythroblasts. Differential interference contrast (DIC) and immunofluorescent micrographs of wild-type (WT) and 4.1R-null enucleating erythroblasts, including nascent reticulocyte (R) and extruding nucleus (N), probed with fluorescein isothiocyanate–conjugated TER 119, specific for GPA (green), or Alexa Fluor 555–labeled rabbit anti–mouse GPC antibody (red). Nuclei were identified by Syto-17 staining (blue). The number of enucleating erythroblasts examined under each staining condition was 6 or more. The images were observed by the use of a Zeiss LSM META 510 Confocal microscope (Carl Zeiss Microimaging Inc) with an APOCHROMAT 63×/1.4 oil DIC objective and acquired by the use of Zeiss Laser Scanning Microscope LSM 510, Version 3.2 SP2 software with a Zeiss AxioCam HRm Rev. 2/3.3V camera. The images were processed with the use of Adobe Photoshop (Adobe Systems Inc).

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