Figure 8. The myeloid CX3CL1/CX3CR1 axis is augmented in Vhl-deficient tumors.

(A) Quantification of soluble CX3CL1 in Vhl WT.2, Vhl-KO.7, and Vhl CX3CL1-DKO CM by ELISA. Data represent technical replicates from 2 independent experiments and were normalized per 10⁶ cells. (B) Relative monocyte migration stimulated by either SF media or CM from Vhl WT.2, Vhl-KO.7, or Vhl Cx3cl1-DKO cells. (C) Average growth curve of all Vhl-KO.7 (gray) and Vhl Cx3cl1-DKO (purple) tumors represented as tumor volume (mm³). Smaller graphs represent biological replicates. (D) Quantification of CD45⁺, CD11b⁺, and CD3⁺ immune infiltrate from Vhl WT, Vhl-KO, and Vhl Cx3cl1-DKO tumors. (E) Quantification of overall TAM, TAM1, and TAM2 infiltration as the percentage of viable cells in Vhl-KO and Vhl Cx3cl1-DKO tumors (F) Protein MFI quantification and representative histogram of CD11c and (G) CD206 in overall TAMs from Vhl WT.2, Vhl-KO.7, and Vhl Cx3cl1-DKO tumors. (H) Percentage of Phrodo⁺ cells as a fraction of viable CD45⁺CD11b⁺F4/80⁺ cells in Vhl WT.2 and Vhl-KO.7, and Vhl Cx3cl1-DKO tumors. (I) Ranked gene expression scores for CX3CL1 and (J) CX3CR1 across 30 nonlymphoid solid tumors queried in TCGA. Data represent biological replicates. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001, by 1-way ANOVA with Bonferroni’s multiple-comparison test (A, B, D, F, and G), 2-way ANOVA with Šidák’s multiple-comparison test (C), and 2-tailed Student’s t test (E and H). Graphs show the mean ± SEM.