Fig 1: Functional investigation of differentially expressed genes in ?d[2] versus ?d[T2] cells(A) IL-9 was measured in supernatants harvested on day 15 from ?d[2] and ?d[T2] cells (values for individual donors are linked by a line; unpaired 2-tailed Student’s t test).(B) IL-9 receptor expression was quantified on ?d[2] and ?d[T2] cells on day 15 using flow cytometry (unpaired 2-tailed Student’s t test).(C) PTGER2 protein expression in ?d[2] and ?d[T2] cells (n = 3 independent donors) was determined by western blotting under reducing conditions. Quantification of band pixel density was performed using ImageJ. Data were normalized for each ?d[T2] cell sample with respect to ?d[2] cells expanded from the same donor.(D) CD300a was quantified on day 15 from ?d[2] and ?d[T2] cells using flow cytometry (unpaired Student’s t test).(E) The indicated ffLuc-expressing tumor cell lines were co-cultivated with ?d[2] and ?d[2-ITGAE] cells for 24 h at 1:1 E:T ratio. Tumor cell viability is shown in comparison with tumor cells that had been cultured alone (means ± SDs; 1-way ANOVA).(F) Supernatants were collected from co-cultures described in (E) after 72 h and analyzed for IFN-? (means ± SDs; unpaired Student’s t test).(G) Effect of exogenous PGE2 on cytotoxicity (means ± SEMs, n = 10 from 5 independent donors) by ?d[2] and ?d[T2] cells when co-cultivated with U937 and KG-1 cell lines for 72 h at 1:1 E:T ratio. Data were normalized with respect to cytotoxicity by ?d[2] and ?d[T2] cells in the absence of added PGE2 (2-way ANOVA). NS, not significant.(H) Effect of exogenous PGE2 on IFN-? production (means ± SEMs, n = 8 from 4 independent donors) by co-cultures described in (G). Data were normalized with respect to IFN-? production by ?d[2] and ?d[T2] cells in the absence of added PGE2 (2-way ANOVA).“All” in (E) and (F) refers to pooled data from solid tumor and leukemic cell lines, respectively.