Figure 8. The effects of Vactosertib on tumor extrinsic MDSC cell profile.

BALB/c mice were inoculated with 1×10⁶ K7M2-Luc (i.v).on Day 0, and then treated with vehicle (p.o) or Vactosertib (50 mg/kg, p.o. 5 days/week) starting on day 28 (4 weeks). 10 weeks after tumor injection, lung samples were collected and examined for expression of CD45.2, CD11b, CD11c, F4/80, Ly6C and Ly6G by flow cytometry. Unbiased immune cell profiling on 2487 live CD45.2+ cells by tDistributed Stochastic Neighbor Embedding (t-SNE) analysis was performed. A) tSNE heatmap plots show the MFI expression level and the distribution of each indicated marker; low (blue) to high (red). B) tSNE density plots of CD45.2+ cells in vehicle or Vactosertib treated samples. C) tSNE dotplot overlays of clusters with significant differences (p<0.05) between vehicle and Vactosertib treatment groups. D) Heatmap showing the normalized MFI of the markers by cluster (low to high; blue to red); arrows indicate significant difference of events between vehicle and Vactosertib groups I. groups E) Bar graphs showing selected clusters with significant difference of events between vehicle and Vactosertib groups (p<0.01). F) The frequency of MDSC cell markers by conventional FACS analysis. Vehicle n=4, Vactosertib n=7 *p<0.05, using an unpaired two-tailed t-test.