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Fig. 2 | Journal of Neuroinflammation

Fig. 2

From: Functional characteristics of Th1, Th17, and ex-Th17 cells in EAE revealed by intravital two-photon microscopy

Fig. 2

Encephalitogenic Th17 cells acquire an ex-Th17 IFN-γ-producing phenotype in vivo. a Experimental setup for TPLSM experiments. Naïve T cells from 2d2.RFP or IL-17 reporter mice were skewed to Th1 or Th17 cells. Flow cytometry was performed to verify appropriate cytokine profile. Staining was performed using antibody staining of PFA-fixed cells. Cells were then transferred to recipient mice. b Representative snapshots from TPLSM videos of different conditions. (i) transfer of 10 million 2d2 Th17-skewed cells, (ii) transfer of 10 million 2d2 Th1-skewed cells, (iii) transfer of 10 million IL-17-reporter Th17-skewed cells, (iv) transfer of 10 million IL-17-reporter Th1-skewed cells. IL-17 reporter mice express eGFP as a marker of Il17a activity, a MOG-specific T cell receptor, and ubiquitously express tdRFP. 2d2.RFP mice express a MOG-specific T-cell receptor and ubiquitously express tdRFP. c Quantification of cells per tissue volume (106 μm3) in videos from different conditions described in (b). Data from at least 3 different mice per condition shown. d Quantification of single cell, living CD4+ cells after either Th1 or Th17 application. Animal scores were above 2. *p < 0.05, **p < 0.01 n = 4–7 animals per group. e Representative disease of 14 mice from two independent experiments. IFNγ expression (right) during Th17-induced disease (left). Expression levels of IFNγ in ex-Th17 were measured when mice displayed clinical scores of 1, 2, and 3. Expression levels have already reached their final level at the onset of disease and remain stable over its course. n = 3

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