Alterations in sialic-acid O-acetylation glycoforms during murine erythrocyte development

Vinay S. Mahajan; Faisal Alsufyani; Hamid Mattoo; Ian Rosenberg; Shiv Pillai

Erythrocyte precursors in the bone marrow can be identified based on CD71 and TER-119 staining and are divided into the Ery-A (TER-119 + CD71 + FSC hi ), Ery-B (TER-119 + CD71 + FSC lo ) and Ery-C (TER-119 + CD71 -FSC lo ) flow cytometric fractions based on CD71, TER-119 and forward scatter (FSC) (Fig 3B) (Chen et al., 2009; Koulnis et al., 2011) . We found that surface 9-O-acetyl sialic acid identified using CHE-FcD staining appears only at the most mature Ery-C stage in the erythroid lineage. We did not observe any CD71 positive cells that lacked TER-119 in the wild-type mouse bone marrow, suggesting that CD71 and FSC alone could be used to identify the Ery-A and Ery-B erythroblast fractions in the bone marrow of Casd1-deficient mice (that lack the TER-119 epitope) ( Fig 3B) . As in wild-type mice, in the bone marrow of Casd1-deficient mice, there was also no detectable surface 9-O-acetylation on CD71 + Ery-A and Ery-B erythroid progenitors based upon staining with CHE-FcD ( Fig 3B) . Our flow cytometry analysis suggests that Casd1-deficient mice exhibit a pan-hematopoietic loss of 9-O-. CC-BY-NC-ND 4.0 International license is made available under a The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It . https://doi.org/10.1101/469254 doi: bioRxiv preprint acetyl sialic acids in the myeloid, lymphoid and erythroid lineages.