Supplementary Figure 8: PU.1 is not essential for neutrophil priming.

a, Distribution of the neutrophil signature (Fig. 6c–h) in different CRISP-seq infected mice. Mice are colored by whether PU.1 was included in the virus mix. The central mark in box plot is median, with 5/95 percentiles at the whiskers and 25/75 percentiles at the box. n = 30 independent animals. b, FACS plots of CRISP-seq output 11 days after transplantation with a mix of LSK cells infected separately with mCherry PU.1 gRNA and BFP control gRNA (mix 3 and mix 4 in Supplementary Table 3). PU.1 KO Ly6G⁺ lack CD11b expression. These results were repeated in four independent animals. c, Gene expression profiles of 6,529 cells pooled from four mice transplanted with a mix of mCherry PU.1 gRNA and BFP control gRNA and grouped by MetaCell analysis. Lower panel indicate enrichments across different samples. d, Defining the mature neutrophil transcriptional program. Differential expression of genes seen solely in a subset of control cells and not found in PU.1 infected cells, compared to the rest of control and PU.1 infected cells (Supplementary Table 2). e, MetaCell analysis of 1,501 LSK cells infected with mCherry PU.1 gRNA and BFP control gRNA (mix 3 and mix 4 in Supplementary Table 3) after stimulation in vitro with G-CSF or GM-CSF. Lower panel indicates enrichments in different samples. f, FACS analyzed cell counts from the in vitro assay after 9 and 14 days. g, Fraction of monocytes in control and PU.1 gRNA infected cells grown with GM-CSF. h, Pooled expression of genes defining the monocyte score (Fig. 6d–j) in cells infected with PU.1 gRNA (y axis) and control gRNA (x axis). i, Representative May-Grünewald Giemsa stain of day 11 in vivo PU.1 KO Ly6G⁺ donor cells highlighting morphological segmentation. Image is representative of at least 2 independent experiments.