Skip to main content
. 2015 Oct 8;5:14896. doi: 10.1038/srep14896

Figure 2. Long-term growth arrest of cancer cells mediated by TR-CD4.

Figure 2

(A) SK37 was co-cultured with or without TR-CD4 or NTR-CD4 for 5 days. Non-adherent T cells were removed by repeated rinses using culture medium and adherent SK37 were further cultured for 10 days. Cell numbers were determined by trypan blue exclusion assays. Fold expansion was calculated by dividing the cell number on day 10 or 15 by the cell number on day 5. (B) SK37 was cultured with recombinant IFN-γ and/or TNF-α for 5 days. Adherent SK37 were rinsed and cultured in the medium without cytokines for 10 days. Cell numbers were determined by trypan blue exclusion assays. (C) SK37 was co-cultured with TR-CD4 in the presence or absence of neutralizing antibodies against IFN-γ (αIFN) or TNF-α (αTNF), or inhibitors for perforin (concanamycin A: CMA) or granzyme B (benzyloxycarbonyl-Ala-Ala-Asp-chloromethylketone: Z-AAD) for 3 days. Non-adherent T cells were removed by repeated rinses and adherent SK37 were further cultured. Cell numbers were determined at the indicated days after the culture. **P < 0.01 at day 9 by Student’s t-test. (D) SK37 was co-cultured with TR-CD4 in the presence or absence of αIFN and αTNF. At day 3 of co-culture, cell cycle of SK37 was analyzed by staining with anti-BrdU antibody and 7-AAD.