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. 2019 Sep 30;9:13991. doi: 10.1038/s41598-019-50614-1

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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IL-12 signaling promotes Bcl-6, IL-21, and ICOS expression in T_FH1-like cells. (A) qRT-PCR to assess expression of the indicated genes in T_FH1-like cells cultured with (teal bars) or without (white bars) IL-12. The data were normalized to Rps18 and presented as fold change relative to T_FH1-like cells cultured with IL-12 (mean of n = 3 ± s.e.m.). (B) Immunoblot analysis of Bcl-6 protein expression in T_FH1-like cells cultured with or without IL-12. Shown is a representative blot of three independent experiments. β-actin was used as a loading control. (C) qRT-PCR to assess expression of the indicated genes in T_FH1-like cells cultured with (blue bars) or without (white bars) IL-12. The data were normalized to Rps18 and presented as fold change relative to T_FH1-like cells cultured with IL-12 (mean of n = 3 ± s.e.m.). (D) Flow cytometry analysis of intracellular expression of IL-21 and IFN-γ in T_FH1-like cells cultured with or without IL-12. Shown is representative data from four independent experiments. (E) The percent of IFN-γ⁺IL-21⁺ cells as assessed by flow cytometry analysis in ‘D’ (mean of n = 4 ± s.e.m.). (F) ELISA analysis of IgM production by B cells co-cultured with the indicated T_FH1-like population at a 3:1 B/T cell ratio for 5 days (mean OD of n = 3 ± s.e.m.). **P < 0.01, ***P < 0.001; unpaired Student’s t-test.