Fig. 5

Inhibition of METTL3 activity induces epigenetic reprogramming of differentiated Treg cells. a Left: Representative dot plots showing the proportion of CD4⁺ T cells treated with either scramble siRNA or METTL3 siRNA while cultured under Treg differentiation conditions for 5 days; right: quantification of CD4⁺CD25⁺Foxp3⁺ Treg cells, n = 4. b Left: representative dot plots showing the composition of CD4⁺ T cells in the STM2457-treated and DMSO control groups under Treg differentiation conditions for 5 days; right: quantification of Treg cells, n = 5. c Volcano plot of differentially expressed genes in the STM2457-and DMSO-treated CD4⁺ T cells isolated from human peripheral blood and cultured under Treg differentiation conditions for 5 days. |log₂foldchange| > 1, p < 0.05; red, upregulated genes; blue, downregulated genes; four biological replicates for each group were used for analysis. d, e KEGG pathway enrichment analysis of upregulated (d) and downregulated (e) genes in STM2457-treated CD4⁺ T cells compared with control cells. f Clustered heatmap of 30 Treg signature genes altered after the inhibition of METTL3. g RT-qPCR analysis validating the differentially expressed Treg signature gene transcripts identified by RNA-seq, n = 3. (*p < 0.05, **p < 0.01, unpaired two-tailed Student’s t test for a and b, one-way ANOVA with Dunnett’s multiple comparisons test for g)