Figure 3.

The levels of Yes‐associated protein 1 (YAP1) and phosphatase and tensin homologue (PTEN) in colorectal cancer (CRC) ‐derived cells and their associations with CRC‐mediated myeloid‐derived suppressor cell (MDSC) induction. (a) Western blot analysis showing the expression of YAP1 and PTEN in the CRC cell lines LS174T, LOVO, SW620 and SW480. (b) CD33⁺ cells were isolated from healthy peripheral blood mononuclear cells using human CD33 MicroBeads and were co‐cultured with the CRC cell lines LS174T, LOVO, SW620 or SW480. Gating strategy for the assessment of the MDSC population used flow cytometry. HLA‐DR⁻ cells were gated from live cells, and CD33⁺ CD11b⁺ cells were further gated as MDSCs. The representative dot plots show the percentages of HLA‐DR⁻ CD33⁺ CD11b⁺ MDSCs induced from CD33⁺ cells by co‐culture with LS174T, LOVO, SW620 or SW480 cells in a Transwell system for 48 hr. The CD33⁺ cells in medium alone were included as a control. (c) FACS density plots and statistical analysis showed that the CRC‐induced MDSCs displayed stronger suppression against T cells, including CD4⁺ and CD8⁺ cells. Representative FACS density plots from one of five experiments are shown. *P < 0·05 compared with the control treatment.