MYO1F deficiency promoted tumor growth by inducing the expansion of TANs. (A) Left: WT and Myo1f−/− C57BL/6 mice were subcutaneously injected with 2 × 105 B16F10 cells, tumor collection at day 21 (N = 5 in each group). Right: The corresponding tumor weight. (B) Tumor growth curve over time of WT and Myo1f−/− C57BL/6 mice were subcutaneously injected with 2 × 105 B16F10 cells. (C) Kaplan–Meier survival curves of B16F10 tumor models described in A (N = 10 in each group). (D) FACS analyses of intratumoral CD45+, CD4+, CD8+ (IFN-γ+ and GZMB+) cells, and neutrophils from tumor tissues described in A. (E) FACS analyses of intratumoral neutrophils from tumor tissues described in A. (F) Immunofluorescence staining of CD11b (green) and CD33 (red) in human melanoma (normal, N = 5; melanoma, N = 45). Scale bar: 200 µm. (G) Quantity statistics of colocalization in immunofluorescence staining (F). (H) Schematic of intratumoral neutrophil transfer from donor (CD45.1 B16F10 tumor-bearing WT or KO mice) to recipient (CD45.2 WT mice). (I) Tumor neutrophil transfer effect on tumor progression. Sorted neutrophils from donor tumor tissues of B16F10-bearing mice transfer into B16F10 tumor models of recipient. Left: Tumor collection on day 24 after subcutaneously injected with 2 × 105 B16F10 cells. Right: Tumor growth curve over time (N = 5 in each group). (J) SX-682 effect on B16F10 tumor models. Left: Tumor collection on day 26 after subcutaneously injected with 2 × 105 B16F10 cells. Right: Tumor growth curve over time (N = 5 in each group). Dose: 50 mg/kg, i.g., bid. (K) FACS analyses of intratumoral CD8+ (GZMB+) T cells (N = 5). (L) Top: Flow cytometry analyses of Ly6G and CD11b in BM CD45+ myeloid cells from WT and Myo1f−/− B16F10 models of day 21. Bottom: Statistic of neutrophil proportion and numbers by counting through FACS (N = 5). (M) Top: Flow cytometry analyses of Ki-67 in CD11b+Ly6G+ (Neu) clusters from BM of WT and Myo1f−/− B16F10 tumor models of day 21. Bottom: Statistic of Ki-67 proportion (N = 5). (N) Left: Representative immunofluorescence staining of CD11b (green) and Ly6G (red) in BM from WT and Myo1f−/− B16F10 model. Right: Statistic of neutrophil numbers by counting colocalization (N = 3). Scale bar: 500 µm. (O) BM neutrophil transfer effect on tumor growth curve over time. Sorted BM neutrophils from donor tumor tissues of B16F10-bearing mice of day 21 were transferred into B16F10 tumor models of recipient (N = 5). (P) FACS analyses of intratumoral CD8+ (GZMB+) T cells (N = 5). Data in A–E, H–M, O, and P represent one experiment of three independent repeats; F and N represent one experiment of two independent repeats. Data are presented as mean ± SD. P values were analyzed by one-way ANOVA test (B, I, J, and O); log-rank (Mantel–Cox) (C); two-tailed unpaired Student’s t test (A, D, E, G, K–N, and P), *P < 0.05, **P < 0.01, and ***P < 0.001. ns, no significance. Neu, neutrophil.
Figure 2.

MYO1F deficiency promoted tumor growth by inducing the expansion of TANs. (A) Left: WT and Myo1f−/− C57BL/6 mice were subcutaneously injected with 2 × 105 B16F10 cells, tumor collection at day 21 (N = 5 in each group). Right: The corresponding tumor weight. (B) Tumor growth curve over time of WT and Myo1f−/− C57BL/6 mice were subcutaneously injected with 2 × 105 B16F10 cells. (C) Kaplan–Meier survival curves of B16F10 tumor models described in A (N = 10 in each group). (D) FACS analyses of intratumoral CD45+, CD4+, CD8+ (IFN-γ+ and GZMB+) cells, and neutrophils from tumor tissues described in A. (E) FACS analyses of intratumoral neutrophils from tumor tissues described in A. (F) Immunofluorescence staining of CD11b (green) and CD33 (red) in human melanoma (normal, N = 5; melanoma, N = 45). Scale bar: 200 µm. (G) Quantity statistics of colocalization in immunofluorescence staining (F). (H) Schematic of intratumoral neutrophil transfer from donor (CD45.1 B16F10 tumor-bearing WT or KO mice) to recipient (CD45.2 WT mice). (I) Tumor neutrophil transfer effect on tumor progression. Sorted neutrophils from donor tumor tissues of B16F10-bearing mice transfer into B16F10 tumor models of recipient. Left: Tumor collection on day 24 after subcutaneously injected with 2 × 105 B16F10 cells. Right: Tumor growth curve over time (N = 5 in each group). (J) SX-682 effect on B16F10 tumor models. Left: Tumor collection on day 26 after subcutaneously injected with 2 × 105 B16F10 cells. Right: Tumor growth curve over time (N = 5 in each group). Dose: 50 mg/kg, i.g., bid. (K) FACS analyses of intratumoral CD8+ (GZMB+) T cells (N = 5). (L) Top: Flow cytometry analyses of Ly6G and CD11b in BM CD45+ myeloid cells from WT and Myo1f−/− B16F10 models of day 21. Bottom: Statistic of neutrophil proportion and numbers by counting through FACS (N = 5). (M) Top: Flow cytometry analyses of Ki-67 in CD11b+Ly6G+ (Neu) clusters from BM of WT and Myo1f−/− B16F10 tumor models of day 21. Bottom: Statistic of Ki-67 proportion (N = 5). (N) Left: Representative immunofluorescence staining of CD11b (green) and Ly6G (red) in BM from WT and Myo1f−/− B16F10 model. Right: Statistic of neutrophil numbers by counting colocalization (N = 3). Scale bar: 500 µm. (O) BM neutrophil transfer effect on tumor growth curve over time. Sorted BM neutrophils from donor tumor tissues of B16F10-bearing mice of day 21 were transferred into B16F10 tumor models of recipient (N = 5). (P) FACS analyses of intratumoral CD8+ (GZMB+) T cells (N = 5). Data in A–E, H–M, O, and P represent one experiment of three independent repeats; F and N represent one experiment of two independent repeats. Data are presented as mean ± SD. P values were analyzed by one-way ANOVA test (B, I, J, and O); log-rank (Mantel–Cox) (C); two-tailed unpaired Student’s t test (A, D, E, G, K–N, and P), *P < 0.05, **P < 0.01, and ***P < 0.001. ns, no significance. Neu, neutrophil.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal