Fig. 4
Effects of IPG0521m treatment on the phenotypes of TI-Tregs. A Distribution of Treg cells. B Expression of Ccr8 by UMAP plot. C UMAP projection of Treg cells, showing the formation of 3 subclusters (Foxp3hi, Foxp3int and Foxp3low). Each dot represents an individual cell, colored according to cell cluster number. D The bubble plot shows the enrichment of distinct markers and Treg-related signature genes of different Treg subclusters. The bubble size represents the percentage of genes in the gene signature (x-axis) expressed in the corresponding cell subclusters (y-axis), and the color bar represents the average expression of genes. E The heat map showing the expression of immunosuppressive genes in Treg cell populations. F The cell proportion of Treg subclusters with or without IPG0521m administration based on the analysis of t-test. G Heat map showing the enrichment of pathways in Treg subclusters. H-I Tregs isolated from H22 liver cancer tissues were treated with mCCL1 (100 ng/ml) and IPG0521m (10 ug/ml) as indicated, and the expression of Lag3 and Ctla4 was evaluated using real-time PCR. J CFSE labeled CD8+ T cells isolated from spleens of healthy mice were co-cultured with Tregs isolated from H22 liver cancer tissues in the presence or absence of mCCL1 (100 ng/ml) and IPG0521m (10 ug/ml), and the proliferation of CD8+ T cells was assessed by means of FCM. K Quantification of CD8+ T cells proliferation described in (J). Data were shown as mean ± SD. *p < 0.05, ***p < 0.001