Fig. 1: Analysis of the LLC tumor microenvironment under SULT2B1b perturbation.

A One representative confocal microscopy imaging of CD3⁺ T cells infiltrating LLC-Mock and LLC-SULT2B1b tumors. Red, CD3⁺ T cells; Blue, DAPI (bars = 200 µm). B Absolute T cell number/field evaluated by counting 4 different experiments with FIJI software (extension of ImageJ) and classical cell counter. DAPI⁺ events (blue) displaying CD3 staining (red) were considered as positive T cells. ***P < 0.001 (Student’s t-test). C Flow cytometry analysis of the percentage of CD45⁺CD3⁺ T cells infiltrating LLC-Mock and LLC-SULT2B1b tumors. *P < 0.05 (Student’s t-test). Mean and s.d. of four experiments. D Flow cytometry analysis of the percentage of CD3⁺CD8⁺INF-γ⁺ T cells infiltrating LLC-Mock and LLC-SULT2B1b tumors. **P < 0.01 (Student’s t-test). Mean and s.d. of two experiments. E Flow cytometry analysis of the percentage of CD3⁺CD4⁺INF-γ⁺ T cells infiltrating LLC^-Mock and LLC-SULT2B1b tumors. P = 0.01 (Student’s t-test). Mean and s.d. of two experiments. (F) LLC-Mock and LLC-SULT2B1b tumor growth. Mean and s.d. of one experiment (n = 10 mice/group). ****P < 0.0001 (Student’s t-test). (G and H) Representative confocal microscopy imaging of CD3⁺ T cells and CD11c⁺ cells (G) and CD3⁺ T cells, CD11b⁺ and CD11c⁺ cells (H) infiltrating LLC-Mock and LLC-SULT2B1b tumors. Red, CD3⁺ T cells; Blue, DAPI, Green, CD11c; Gray, CD11b (bars = 200 µm). (I) Quantification of the fields of DC/T cell clusters evaluated by counting 4 different experiments with FIJI software (extension of ImageJ) and classical cell counter. The number of sections in each image displaying the presence of both DCs (CD11c⁺CD11b⁺) and T cells (CD3⁺) was considered as a positive DC/T cell cluster. *P < 0.05 (Student’s t-test).