Nbea deficiency affects vesicle trafficking components at the transcriptional level. (A-B) Control and Nbea-shRNA–treated progenitors were transplanted into lethally irradiated recipients. mCherry⁺ cells were recovered from transplanted mice 10 weeks after transplant and subjected to bulk RNA sequencing. (A) Experimental schematic. (B) Pathway analysis based on transcriptional differences among control and Nbea-shRNA–treated progenitors in (i) biological processes and (ii) cellular components. Differences related to vesicle trafficking highlighted with an arrow. (C-D) E14.5 FL LT-HSCs were isolated from Nbea^+/+ and Nbea^{–/ –} embryos and subjected to bulk RNA sequencing. (C) Experimental schematic. (D) Pathway analysis including (i) biological processes and (ii) cellular components showing differences among Nbea^+/+ and Nbea^–/– E14.5 FL LT-HSCs. (E) Representative confocal images of BM HSC and NBEA/RCAS1 colocalization analysis (Pearson correlation coefficient). NBEA-A488 shown in green, RCAS1-A647 (Golgi) in red, and nuclei (DAPI) in blue. (F) qPCR analysis of NOTCH-regulated genes in E14.5 FL-HSC in Nbea^+/+ and Nbea^–/–. Data in panel B are from 2 independent samples. Data in panel D are from 1 sample. Data in panel F are from 3 samples, represented as mean ± SEM. ∗P < .05; ∗∗P < .005, relative Nbea^+/+.