Fig. 3: ILC1 TNFα rather than IFNγ suppresses the differentiation of LSCs into CD11b⁺CD206⁺ immunosuppressive, macrophage-like leukemia-supporting cells.

a Human LSCs were co-cultured without or with human HD ILC1s for 3 days. Representative images of Wright-Giemsa staining of cells (20× magnification, scale bar, 100 µm, n = 3). b, c Human LSCs were co-cultured without or with HD ILC1s, HD ILC1s plus anti-IFNγ, or HD ILC1s plus anti-TNFα for 3 days. Representative flow cytometry plots (b) and summary data of the absolute cell number (c; left) and percentage (c; right) of CD11b⁺CD206⁺ cells (n = 7 HDs; n = 7 AML patients). d, e Human LSCs were co-cultured without or with IFNγ or TNFα for 3 days. Representative flow cytometry plots (d) and summary data of the absolute cell number (e; left) and percentage (e; right) of CD11b⁺CD206⁺ cells (n = 4 AML patients). f, g Human LSCs were co-cultured without or with HD ILC1s for 3 days. Total colony-forming cells (f) and colony-forming myeloid progenitors (CFU-GM and CFU-G; g) were counted at each round of plating (n = 3 HDs; n = 3 AML patients). (h) Identification of FLT3-ITD mutation by PCR. A representative 3% agarose gel electrophoresis of PCR products. The 329 bp fragment indicates the wild type. The presence of both a band at 329 bp and a fragment larger than 329 bp (>329) indicates FLT3-ITD mutation⁵⁶ (n = 2 AML patients). Data are representative of two (a, d, e, and h) and three (b, c, f, and g) independent experiments and are presented as the mean ± s.d.; P values were calculated by one-way ANOVA (c and e) and two-tailed unpaired Student’s t test (f and g). For (f), P values were calculated after the Log₁₀ transformation due to large variations. Source data are provided as a Source Data file.