Extended Data Fig. 4: Characterization of splenic immune cell subpopulations by single-cell analysis.
a–d, C57BL/6-hPD1 mice were subcutaneously inoculated with MC38 tumor cells and treated with IgG4-mEry, anti-PD1 or αPD1-mEry twice weekly once tumors reached approximately 100 mm³. Splenocytes were isolated 3 days after the second treatment from one mouse per treatment group and analyzed by mass cytometry (a) and scRNA-seq (b–d).a, Marker heatmap used for immune cell clustering shows the median expression of each marker per cluster, scaled for visualization, as revealed by mass cytometry. b, Combined t-SNE plot of all splenic cells from tumor-bearing mice with different treatments, as revealed by scRNA-seq. c, e, f, Dot plots show scaled expression values of discriminative gene sets per defined cluster. d, scRNA-seq analysis demonstrating sample preference of each cluster, estimated by RO/E values using a two-tailed chi-square test.
