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. 2015 Mar 6;6:6436. doi: 10.1038/ncomms7436

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(a) Geometric mean fluorescence intensity (MFI) of ICOS on naïve, effector CD4⁺ and Tfh cells from Mir146a^+/+: Mir146a^−/− and Mir146a^+/+: Mir146a^+/+ mixed bone marrow chimeras 16 weeks post reconstitution. (b) Percentages of Tfh cells in wild-type mice immunized with sheep red blood cells (SRBC) and treated with different amounts of anti-ICOSL blocking antibody on days 0, 3 and 6 post immunization. Each symbol represents the mean and error bars represent the s.d. of three biological replicates. (c) Percentages of Tfh, GC B and Treg cells in 9-week-old wild-type (+/+) and miR-146a-deficient mice (−/−) treated with 25 μg anti-ICOSL blocking antibody or PBS control on day 0, 3 and 6 following SRBC immunization. (d) Percentages of Tfh, GC B and Treg cells in 9-week-old Mir146a^+/+ Icos^+/+, Mir146a^+/+ Icos^+/−, Mir146a^−/− Icos^+/+, Mir146a^−/− Icos^+/− mice following SRBC immunization. (e,f) Adoptive transfer of purified naïve (CD44^low CD25⁻) CD4⁺ T cells from either Mir146a^+/− Icos^+/+, Mir146a^+/− Icos^+/−, Mir146a^−/− Icos^+/+ or Mir146a^−/− Icos^+/−, together with Mir146a^−/− Icos^+/+ B cells and Mir146a^+/+ Icos^+/+ Tregs into Rag1^−/− recipients according to the strategy shown in e. The recipient mice were immunized with SRBC 14 days after transfer and killed 7 days post immunization. (f) Percentages of Tfh and GC B cells in Rag1^−/− recipient mice following adoptive transfer and SRBC immunization. (g,h) ICOSL MFI on GC B (g, left), non-GC B (g, right) and a mixture of CD11b⁺ and CD11c⁺ cells (h) from SRBC-immunized wild-type (+/+) or miR-146a-deficient (−/−) mice following stimulation with LPS, anti-CD40 antibody or isotype control. Heights of bars represent the mean and error bars represent s.d. of three technical replicates. Each symbol represents one mouse and the horizontal bars represent the median values. Data are representative of at least two independent experiments. Statistical significance was determined using paired Student’s t-test (a), unpaired Student’s t-test (b,g,h) or Mann–Whitney U-test (c,d,f). NS, not significant, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.