Figure 4. Exhausted T cells exhibit motile features.

(A) Volcano plots of differentially expressed genes between CD38^hi (top 33rd percentile of all samples) and CD38^lo (bottom 33rd percentile) CRC samples. Red dots are above the cutoff for P = 0.05. (B) Gene ontology analysis of the upregulated genes in the CD38^hi versus CD38^lo samples. (C) Fold-change of promotility genes in exhausted versus nonexhausted T cells from RNA-Seq of melanoma or lung cancer samples as reported in refs. 14–17. (D) Schematic diagram of the experimental design for E–G. (E) Representative images of PD-1 staining for RFP (d14) and GFP in OT-I cells (d4) that resided in the same field. Scale bars: 30 μm. (F) Speed-based, color-coded track displacement of GFP and RFP OT-I cells. (G) PD-1 intensity on OT-I cells was plotted against the mean speed of each cell. Data were pooled from 3 different ROIs and are representative of 2 independent experiments. P and r² values were obtained by linear regression analysis.