MYO1F-deficient neutrophil suppressed immune response during tumor progression . (A) Cell migration ability evaluated by chemotaxis transwell induced by CXCL2; passed cells adhered to the lower chamber surface were counted. (B) Gate strategy of BM neutrophils from B16F10 tumor model at day 21. (C) FACS analyses of Ki-67 in CD45⁺Ly6G⁻ clusters from BM of WT and Myo1f^−/− B16F10 tumor models at day 21 (N = 5). (D) FACS analyses of neutrophils in peripheral blood and spleen from B16F10 tumor model at day 21 (N = 3). (E) Schematic of BM neutrophils transfer from donor (CD45.1 B16F10 tumor-bearing WT or KO mice) to recipient (CD45.2 WT mice). (F) Gate strategy of BM neutrophils sorting from B16F10 tumor model at day 21. (G) Tumor collection at day 20 after tumor inoculation. (H) Top: Tumor growth curve over time. Bottom: Tumor collection at day 21 after tumor inoculation (N = 10). (I) Kaplan–Meier survival curves (N = 20). (J) FACS analyses of intratumoral CD11b⁺Ly6G⁺ neutrophils from tumor tissues (N = 5). (K) FACS analyses of intratumoral CD8⁺ (IFN-γ⁺ and Ki-67⁺) cells from tumor tissues (N = 5). (L) Left: Immunofluorescence staining of CD11b (green) and Ly6G (red) in BM from B16F10 models. Right: Statistic of numbers by counting colocalization (N = 3). Scale bar: 200 µm. (M) Counts of total CD11b⁺Ly6G⁺ neutrophils in one tibia by FACS (N = 5). (N) Gene set enrichment analysis of inflammatory response signaling pathway and immune response signaling pathway. Data in A–J and L represent one experiment of three independent repeats; K and M represent one experiment of two independent repeats. Data are presented as mean ± SD. P values were analyzed by one-way ANOVA test (H); log-rank (Mantel–Cox) (I); two-tailed unpaired Student’s t test (A, C, D, and J–M), *P < 0.05, **P < 0.01, and ***P < 0.001. ns, no significance; Neu, neutrophil; TB, tumor bearing.