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. 2007 Nov 21;2(11):e1204. doi: 10.1371/journal.pone.0001204

Figure 6. Functional consequences of H-RasG12V acquisition.

Figure 6

Purified CD56+ NK or CD3+ T cells were co-cultured for 24 hours with either B721 or HEK293 transfectants as indicated. The supernatants were collected and levels of IL-2, IL-4, IL-5, IL-10, IFN-γ and TNF-α were determined using the Th1/Th2 CBA kit (∼300 beads collected per analyte). (a) Overlay of the two dot plots obtained from supernatants collected from either NK/B721-GFP or NK/B721-G12V co-cultures. Numbers represent the MFI of the respective cytokine. (b) The raw data was computed with the BD™ CBA Software to calculate the levels (pg/ml) of IFN- γ in the indicated supernatants. (c–d) NK and T cells were compared for cytokine IFN- γ (c) or TNF-α (d) secretion in response to either B721-GFP or B721-G12V. (e) Overlay of the two dot plots from supernatants collected from either NK/HEK293-tH or NK/HEK293-G12V co-cultures. (f) The raw data was computed with the BD™ CBA Software to calculate the levels (pg/ml) of IFN- γ in the indicated supernatants. Bars represent the mean±SD of triplicates from a typical experiment out >5 performed. (g) Freshly isolated PBLs were labeled with CFSE and co-cultured with irradiated B721−GFP or B721−H-RasG12V (modified MLR). After 7 days, viable T lymphocytes in the co-cultures were analyzed by FACS for CFSE-dilution (% proliferation). Dots linked by lines represent results from the same donor. *, P = 0.001; **, P = 0.01; ***, P<0.001 (n = 300), and ****P = 0.02 (n = 6).