Extended Data Fig. 3: Virma expression is essential for m6A modification.
Deletion efficiency of floxed Wtap alleles in splenic T cells showing flow cytometry with Wtap-specific antibodies on subsets gating on CD44 and CD62L in splenic CD4+ T cells. (n = 6, three independent experiments) b, Surface staining of TCRβ on subsets gating on CD44 and CD62L in splenic CD4+ T and CD8+ T cells. (n = 6, three independent experiments). c, Percentage of the different congenic marked hematopoietic CD45+ cells (c) and flow cytometry analysis of splenocytes (d) from bone marrow chimeras harbouring WT (CD45.1+) and Wtapfl/fl; Cd4-Cre (CD45.2+) hematopoietic cells, assessed for expression of CD4 and CD8 after identifying CD45.1 or CD45.2 congenic cells. Numbers adjacent to outlined areas indicate the percentages of cells in each gate. (n = 3 individual mice in one experiment). e, Surface staining of CD44 and CD62L on CD4+ and CD8+ T cells after gating on CD45.1 or CD45.2 congenic splenocytes of the bone marrow chimeras. Numbers adjacent to outlined areas indicate percentage of cells in each gate. (n = 3 individual mice in one experiment). f, Immunoblot analysis of Virma protein in extracts from MEF cells showing Gapdh as a loading control. (n = 3, two independent experiments). g, m6A abundance as determined by LC/MS/MS analysis in the oligo-dT-purified mRNAs of Virma-depleted and Cre expressing MEF cells. (n = 3 biological replicates), (mean ± s.e.m, unpaired two-tailed t-test). h, Stainings to identify CD4+CD25+Foxp3+ Treg cells among thymocytes of Virmafl/fl or Virmafl/fl; Cd4-Cre mice. Numbers adjacent to outlined areas indicate the percentages of cells in each gate. (WT, n = 5; KO, n = 4, two independent experiments).
