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. 2018 Jun 28;96(9):994–1007. doi: 10.1111/imcb.12164

Figure 2.

Figure 2

KN6 γδ T‐cells supplemented with IL‐1β, IL‐21 and IL‐23 differentiate toward γδ17 cells in cultures supplemented with Notch1 Ligand Dll4. (a) Day 4 cellularity of KN6 cells on the indicated stromal cells and supplemented with cytokines as shown. (b) Day 4 flow cytometric analysis of γδ17 associated surface markers CD44 and CD62L. (c, d) KN6 cells were stimulated for 6 h with PMA/Ionomycin +CK and (c) analyzed by flow cytometry for cytokine production by KN6 cells. (d) Flow cytometric analysis for CD27 expression on the indicated KN6 γδ T‐cell cytokine expressing subsets. Data shown represent at least 3 independent experiments. KN6 cells were supplemented with the indicated cytokines and cultured on the indicated stromal lines (+CK denotes addition of IL‐7, IL‐1β, IL‐21, IL‐23 on D0). KN6 cells were pre‐gated for DAPI − CD45+and TCRγδ+. The data and error bars are presented as standard error of the mean (s.e.m.). Statistical significance was determined using a two‐tailed unpaired Student's t‐test as P < 0.05 (**P < 0.01).