Fig. 2: TP63 suppresses CD8⁺ T cells infiltration and activation in immune-competent murine SCC models.

A Schematic graph of the in vivo syngeneic experiments. B TP63 expression assessed by western blotting in murine SCC cell lines transduced with Dox-inducible Trp63 shRNA. The similar results were repeated in three biologically independent cells. C Dotplot showing the expression levels of representative marker genes across each immune cell type. For scRNA-seq experiment, each group included 4 tumors from 4 mice, which were combined and dissociated into single cells capture. A total of 1911 CD45⁺ immune cells were analyzed. D UMAP plots of the clustering of 1911 cells from 854 Scramble and 1,057 Trp63 knockdown cells, showing all the intratumoral immune cells (upper) or the immune cells in either Scramble or shTrp63 MOC22 tumors (bottom). E The proportion of each immune cell type in the Scramble and Trp63 knockdown tumors. F UMAP plots showing the subgroups of 645 CD8⁺ T cells from Scramble and shTrp63 MOC22 tumors. G The expression levels of canonical markers for each cell cluster. H The proportion of CD8⁺ T cell subgroups in Scramble and shTrp63 MOC22 tumors. I The proportion of CD3⁺ and CD8⁺ in CD45⁺ immune cells and CD69⁺, GZMB⁺ and IFNγ⁺ in CD8⁺ T cells from shTrp63 or Scramble HNM007 and AKR allografts revealed by FACS analysis. Bars represent mean ± SD of three biologically independent experiments. For the comparison of each population between the Scramble and shTP63 group in HNM007 and AKR allografts: CD45⁺ CD3⁺ (P = 0.0434/0.0792), CD45⁺ CD8⁺ (P = 0.0072/0.7600), CD8⁺ CD69⁺ (P = 0.0003/0.0033), CD8⁺ GZMB⁺ (P = 0.0229/0.0500), CD8⁺ IFNG⁺ (P = 0.0017/0.0495). P values were determined using a two-sided t-test. *P < 0.05, **P < 0.01, *** P < 0.001. The gating strategy are provided in Supplementary Fig. 3B. Source data and the exact cell number of each subgroup in Fig. 1C and E–H are provided as a Source Data file.