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. 2022 Oct 3;2(10):1104–1118. doi: 10.1158/2767-9764.CRC-22-0117

FIGURE 6.

FIGURE 6

SHP2 facilitates growth factor–induced resistance to T-cell cytotoxicity via regulation of PD-L1. A, Schematic of T-cell cytotoxicity assays. Elements in the scheme were created using BioRender. CD8+ T cells are isolated from the spleens of tumor-bearing mice. The MBC cells are treated with growth factors and inhibitors, and cocultured with CD8+ T cells. The dead cells are quantified by with Incucyte imaging. B, Representative images of the MBC cells treated with FGF2 (20 ng/mL) and PDGF (100 ng/mL) at 1 hour following coculturing with T cells. The MBC cells without T cells served as background. C, Bar graph comparing the percentage of dead cell counts of FGF2 and PDGF groups to the no stimulation (NS) group. *, P < 0.05; n = 4 individual repeats. D, Representative images of the MBC cells treated with FGF2/PDGF and TNO155 (5 μmol/L) at 1 hour following coculture with T cells. E, Bar graph comparing the percentage of dead cell counts with different treatments. *, P < 0.05; ***, P < 0.001, n = 9. Histogram of cell surface PD-L1 using flow cytometry (F) and bar graph (G) comparing fold changes of PD-L1 MFIs in D2.A1 cells induced by PDGF and treated with different inhibitors. *, P < 0.05; **, P < 0.01; ***, P < 0.001; n = 3. H, Histogram of cell surface PD-L1 using flow cytometry in D2.A1 cells induced by 3D culture on a fibronectin-coated scaffold and treated with the indicated inhibitors. I, Bar graph comparing fold change of PD-L1 mRNA of D2.A1 cells cultured and treated as in H. **, P < 0.01; n = 3.