Figure 9.

IDO/Kyn/AhR regulated Treg differentiation in CRC. (A, B) Representative frequency of intratumoral Tregs was detected by flow cytometry (n = 6). (C, D) Annexin V expression on intratumoral Tregs and its mean fluorescence intensity were determined by flow cytometry (n = 6). (E, F) The expression of Ki67 and its mean fluorescence intensity of Tregs were shown (n = 6). (G) The percentage of Tregs in spleen lymphocytes was analyzed (n = 6). (H) Kyn-to-Trp ratio in the colon section was measured by enzyme-linked immunosorbent assay (n = 6). (I, J) The expressions of cytoplastic protein AhR and nuclear protein AhR in TIL were analyzed by Western blot (n = 6). (K) Expression of FoxP3 was detected by immunochemical staining of paraffin-embedded colon sections (scale bar = 100 μm). (L) Expression of FoxP3 was detected by immunofluorescent staining of paraffin-embedded colon sections (scale bar = 50 μm, 10 μm). (M) Expression of AhR was analyzed by immunofluorescence cytochemistry (scale bar = 50 μm, 10 μm). The results are representative of at least 6 independent experiments and expressed as mean ± SD. ∗P < .05, ∗∗P < .01 indicates significant difference.