Ikzf1 promotes terminal differentiation of mouse fetal liver derived erythroid cells

Abstract

Objective To explore potential function of IKAROS family zinc finger 1 (Ikzf1) in terminal erythroid differentiation. Methods Single-cell regulatory network inference and clustering (SCENIC) was used to predict transcription factors of terminal erythroid differentiation based on single cell transcriptomes of human and mouse bone marrow erythroid cells. Expression of the predicted Ikzf1 during terminal erythroid differentiation was acquired from public RNA-seq data and validated by RT-qPCR. Short hairpin RNA (shRNA) interference was performed to knock down Ikzf1 in mouse erythroid cells from fetal liver with a GFP cassette to mark successfully transfected cells. The knockdown efficiency in GFP+ cells was verified by RT-qPCR. The effect of Ikzf1 knockdown on erythroid differentiation and enucleation was analyzed by flow cytometry. Results SCENIC predicted that the transcription factor Ikzf1 regulated both human and mouse erythroid differentiation. RNA-seq data and RT-qPCR showed highly expressed Ikzf1 at proerythroblast and basophilic erythroblast stages of terminal erythroid differentiation. Ikzf1 expression was indeed down-regulated by shRNA in erythroid cells from mouse fetal liver. Flow cytometry analysis showed that knockdown of Ikzf1 affected terminal differentiation and significantly reduced the ratio of erythroid enucleation (P＜ 0.05). Conclusions Transcription factor Ikzf1 is highly expressed in early stage erythroblasts and involved in terminal erythroid differentiation as well as enucleation.

Keywords

• ikzf1|terminal erythroid differentiation|enucleation