Atf4 promotes erythroid differentiation by regulating ribosome biogenesis via RPS19BP1
To elucidate the mechanisms underlying the role of Atf4 deficiency in erythropoiesis, we performed 10X Genomics single-cell RNA sequencing on Lin-cKit+ cells from Atf4 hematopoietic cell-specific conditional knockout mice (KO) and Atf4 floxed mice (WT) at steady state (referred to KO_BM and WT_BM) and after secondary transplantation (referred to cKO_t and WT_t). Furthermore, we also sorted CMP cells from Atf4 hematopoietic cell-specific conditional knockout mice (KO) and ATF4 floxed mice (WT) for 10X Genomics scRNA-seq. Overall design: We performed 10X Genomics scRNA-seq of fluorescence-activated cell sorter (FACS)-sorted bone marrow (BM) Lin-cKit+ cells from Atf4 floxed mice (WT) and Mx1cre; Atf4fl/fl (KO) mice at 4 weeks post-deletion. We also enriched Lin-cKit+ cells from the BM of secondary recipient mice for an additional scRNA-seq analysis. In total, we captured 33,237 cells, of which 29,026 passed quality control. To further determine the transcriptional profiles in CMP cells, we performed 10x Genomics scRNA-seq of FACS-sorted CMP cells from WT and KO mice at 4 weeks post-deletion. And 19,706 of 21,914 sequenced CMP cells were kept after quality control.