Figure 3.

The BM Csf2-positive cluster is composed of ILC2s. (A) Histograms of surface markers in GM-CSF⁺ BM cells from 5-FU–treated WT mice. The tinted lines show the background level. Results shown are representative of two independent experiments. (B) qRT-PCR analyses for the expression of Csf2 in total BM cells and Sca1⁺ BM cells from 5-FU–treated mice; n = 3, representative of two independent experiments. Bars indicate expression level relative to that in total BM cells at day 0. (C–E) Single-cell RNA-seq analysis of Sca1⁺ BMMNCs sorted from 5-FU–treated WT mice. (C) t-SNE projection highlighting Csf2 expressing cells (number of detected mRNA molecules >0). The arrow shows a cluster expressing Csf2. (D) t-SNE projection of the expression pattern for Gata3, Il1rl1, Il2ra, and Cd3e. The color of each cell represents the log₁₀(number of molecules per cell). The arrows are in the same position as in C. (E) Six main clusters generated by t-SNE analysis. Each cluster expressed Cd11b, Cd19, Cd34, Cd3e, Flt3, and a combination of Gata3, Il1rl1, and Il2ra. For the 5-FU–treated mice groups, mice were injected intravenously with 200 mg/kg 5-FU 2 d before being euthanized (A–E). Results are shown as mean ± SEM, and each dot represents an individual mouse. Statistical significance was determined by unpaired Student’s t test (B). Coord, t-SNE coordinate.