Fig. 7

Hk3 deficiency and creatine treatment reprogrammed macrophage polarization in vivo and in vitro. A Experimental design. Hk3-ko or WT mice were fed creatine-containing water or not. B, C Hk3-ko and creatine-containing diets promoted tumour growth in subcutaneously injected model mice (B). Boxplot showing the difference of tumour weight among different groups (C) (Data presented as mean ± s.d.; P values were calculated via two-tailed Student’s t test; n = 6). Dots refer to independent samples. D–F Representative flow cytometry analysis of tumour fractions from different groups. Barplot showing the discrepancy in pan TAMs (D, Cd45⁺F4/80⁺Cd11b⁺ cells), M2-like TAMs (E, Cd45⁺F4/80⁺Cd11b⁺Cd206⁺ cells) and M1-like TAMs (F, Cd45⁺F4/80⁺Cd11b⁺Cd86⁺Mhc-II⁺cells) among different groups. Data presented as mean ± s.d.; P values were calculated via two-tailed Student’s t test; n = 3. G, H Effect of Hk3 knockdown (Hk3 sh) and 1 mM creatine pretreatment (+ Cr.) on the chemotactic ability of RAW264.7 cells, with IL-4 for 24 h (H) or not (G), co-cultured with mEC25 cells. Data presented as mean ± s.d.; P values were calculated via two-tailed Student’s t test; n = 5. I Hk3 knockdown (Hk3 sh) and 1 mM creatine pretreatment (+ Cr.) promoted M2-like TAM polarization in RAW264.7 cells. Data presented as mean ± s.d.; P values were calculated via two-tailed Student’s t test; n = 3. See also Fig. S8 and Table S2