Fig. 3

Rigosertib promotes T cell and NK cell responses but attenuates tumor-associated M2 macrophages in the tumor microenvironment. YUMM3.3 tumors were collected 17 days post-treatment. a, h Live CD45⁺ leukocytes were concatenated after downsampling to 20,000 events for subsequent high-dimensional data analysis to normalize the contribution among samples under different treatments. Samples were then analyzed in parallel by t-SNE and manually gated leukocyte populations were overlaid onto the total t-SNE map using FlowJo 10.5.3. b-e, g, i Flow cytometric and f IHC analysis of YUMM3.3 tumors at day 17 post-treatment. j Flow cytometric tSNE analysis of PBMC samples from YUMM3.3 tumor-bearing mice treated with RGS before and after CD4 and/or CD8 antibody depletion treatments. (K) Tumor weight (day 16 post RGS treatment) of YUMM3.3 tumors in C57BL/6 mice with and without CD4 and/or CD8 depletion. a-i pooled data obtained from at least two different experiments (n = 5 ~ 10) are shown. j, k were replicates (n = 10 per group)