Fig. 3: Analysis of T and B cells in tumor and TLS revealed by scRNA-seq.
a The design of collecting 4T1 tumors and TLS is depicted schematically. BALB/c mice (n = 3 per group) were implanted s.c. with 5 × 105 viable 4T1 cells in right flanks, and tumors to reach a palpable size of approximately 5 mm in diameter were peritumorally administered with cisplatin followed by three doses of CR108 in 7-day intervals. Mice received PBS-treated alone were used as the negative control. The cisplatin + CR108 combo-treated 4T1 tumors were surgically separated into TLS nodules (labeled as “TLS”) and external TLS-depleted tumor themselves (labeled as “Tumor”); whereas tumors (labeled as control Tumor) from the PBS group were used as controls. All samples were collected 25 DPI to prepare for next-generation sequencing (NGS), respectively. The image was created with www.Biorender.com and was licensed. b t-distributed stochastic neighbor embedding (t-SNE) plot of immune cells in TLS, with cell counts indicated in parentheses. c The ratio of naïve, costimulatory, Treg, and inhibitory T cells was plotted against each treated group. d RNA velocity analysis for T and NK cells, with the color-coded for pseudotime. e t-SNE plot of T and NK cells, with each color representing one of the subgroups in TIL and TLS. Statistics: The Wilcoxon rank-sum test was performed to compare ratios of each kind of T cell in different treatment groups and resulted in P values were corrected for multiple testing via the method of Benjamini & Hochberg.
