Extended Data Fig. 7: Treatment with FTY720 does not affect the phenotype and proliferation of TCRHh blocked with anti-IL-10R.
a. TCRHh were transferred into Hh colonised host mice, treated with daily FTY720 from day 15 and anti-IL-10R at day 17. b. Frequency of Foxp3 among total TCRHh CD4 T cells (left), geometric MFI of Foxp3 expression in TCRHh Tregs (centre) and geometric MFI of c-MAF expression in TCRHh Tregs (right) in control and anti-IL-10R treated mice at day 20. c. Ki-67 of total CD4 T cells of TCRHh and host cells (left), Ki-67 of TCRHh Tregs (centre) and Ki-67 of TCRHh Th17 cells (right) in control and anti-IL-10R treated mice at day 24 with and without FTY720 blockade. d. Representative FACS plot of c-MAF versus Ki-67 in TCRHh Tregs (left) and c-MAF geometric MFI of Ki-67+ vs Ki-67- TCRHh Tregs (right). e. Geometric MFI of Foxp3 (left) and c-MAF (right) of TCRHh Tregs in control and anti-IL-10R treated mice with and without FTY720 blockade. g. Expression of Ifnγ (left), Il17α (centre) and Il6 (right) in caecum tissue by qPCR in control and anti-IL-10R treated mice at day 24. g. Expression of Il27 (left), Il10 (centre left), Il2 (centre right) and Il23a (right) in caecum tissue by qPCR in control and anti-IL-10R treated mice at day 24. h. Representative H and E staining (left top and bottom) and histopathology score (right top) of caecum tissue from control and anti-IL-10R treated mice at day 24. Scale bar represents 1 mm. i. Colon histopathology score in control and anti-IL-10R treated mice with and without FTY720 blockade. j. Representative immunofluorescent staining of Foxp3 and Ki-67 in LA (left, arrows mark Ki-67 negative Tregs), control LP (centre, arrows mark Ki-67 negative Tregs), and anti-IL-10R treated LP (right, arrows mark Ki-67 positive Tregs). k. Frequency of Ki-67 positive Tregs in the LP of control and anti-IL-10R treated mice (left) and frequency of Ki-67 positive Tregs in the LP and LA in both control and anti-IL-10R treated mice (right). b-i: 6 individual mice per group representative of 2 independent experiments. f: 11 control and 11 anti-IL-10R treated mice combined over 2 independent experiments. J-k: 5 individual mice per group representative of 2 independent experiments. b, d, f, g, k: two-tailed t test. c, e, i: one-way ANOVA using a Tukey’s multiple comparisons test.
