Loss of MLL3/GRHL2 attenuates T cell infiltration in the tumor microenvironment. (A) Schematic illustration of the workflow for the investigation of infiltrated T cell infiltration in syngeneic mouse models. (B and D) FACS plots of Zombie⁻CD45⁺CD3⁺CD4⁻CD8⁺T cells and Zombie⁻CD45⁺CD3⁺CD4⁺CD8⁻ T cells in indicated samples. The left-most panel of B was also presented as the last gating plot (CD8 versus CD4) in Fig. S3 B. (C and E) Bar graph of the percentage of CD8⁺ T cells and CD4⁺ T cells among live cells (n = 5 for each group). (F) IF staining of infiltrated CD8⁺ T cells in the xenograft tumors and bar graph of quantification of CD8⁺ T cell (n = 3 biological replicates). Scale bar = 100 μm. (G) Schematic illustration of the workflow of T cell migration assay using transwell system. Sorted CD8⁺ T cells were cultured for 48 h using conditioned medium harvested from either MLL3 KD or GRHL2 KD cell line. (H and I) The number of migrated CD8⁺ T cells into the bottom counted by hemocytometer (n = 3 biological replicates). *P < 0.05; **P < 0.01; ***P < 0.001.