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. 2021 Feb 26;12:1333. doi: 10.1038/s41467-021-21594-6

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© The Author(s) 2021

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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Fig. 5 — a Graphic representation of the wild-type (Mettl3-WT) and catalytic domain dead (Mettl3-Mut; DPPW to APPA) METTL3 constructs. b Scheme of retrovirus transduction experiment. SMARTA cells were transduced with indicated structures by using a retrovirus transduction system. Then, the transduced cells were adoptively transferred into congenic CD45.2⁺ wild-type mice followed by LCMV-Armstrong infection, and analyzed on day 5 post viral infection. c Flow cytometry analysis of METTL3 gMFI in GFP⁺CD4⁺ SMARTA cells from recipient mice. Quantitation of METTL3 gMFI is shown on the right (n = 3 per group). d Summary of the frequency and cell numbers of retrovirus-transduced SMARTA CD4⁺ T cells in host mice (n = 3 per group). e, f Flow cytometry analysis of CD44⁺CXCR5⁺ T_FH populations and CD44⁺CXCR5^– T_H1 subsets gated on SMARTA GFP⁺CD4⁺ T cells from host mice adoptively transferred with empty vector (EV), Mettl3-WT, or Mettl3-Mut retrovirus-introduced SMARTA cells. Summary of the frequency and cell numbers of T_FH cells and T_H1 cells are shown in f (n = 3 per group). g, h Flow cytometry analysis of PD-1^hiCXCR5⁺ GC T_FH populations gated on SMARTA GFP⁺CD4⁺ T cells from host mice adoptively transferred with EV, Mettl3-WT, or Mettl3-Mut retrovirus-introduced SMARTA cells. Summary of the frequency and cell numbers of GC T_FH cells are shown in h (n = 3 per group). i Quantitative RT-PCR analysis of mRNA abundance of T_FH cell-related genes in CD44⁺CXCR5⁺ T_FH cells ectopically expressed with EV, Mettl3-WT, or Mettl3-Mut, relative expression was normalized to Ctrl cells transduced with EV retrovirus (n = 6 for Ctrl EV, Mettl3^fl/flCd4-Cre-EV, and Mettl3^fl/flCd4-Cre-Mettl3 WT group; n = 5 for Mettl3^fl/flCd4-Cre-Mettl3 Mut group). Data are representative of two independent experiments. Error bars indicate standard error of the mean. P value was calculated by one-way ANOVA, followed by unpaired two-tailed Student’s t test for indicated pairwise comparisons.