Fig. 5: Serine downregulates Supervillin via S-adenosyl-methionine-mediated tri-methylation of H3K9.

a The metabolic flux of ¹³C₃-serine in CD34⁺ cells undergoing MK differentiation, m+0 denotes metabolites without labeled carbon atom, while m+1, m+2, m+3, and m+4 denotes metabolites with 1, 2, 3, and 4 labeled carbon atom. b Schematics of serine metabolism in MKs. c Generation of CD41a⁺CD42b⁺ MKs of CD34⁺ cells with indicated concentrations of methionine (mean ± SD, n = 3 independent experiments). d Generation of CD41a⁺CD42b⁺ PLPs from CD34⁺ cells with indicated concentrations of methionine (mean ± SD, n = 3 independent experiments). e Generation of CD41a⁺CD42b⁺ MKs from CD34⁺ cells infected with scramble or SHMT2-shRNA virus with or without serine treatment (mean ± SD, n = 3 independent experiments). f Generation of CD41a⁺CD42b⁺ PLPs from CD34⁺ cells infected with scramble or SHMT2-shRNA virus with or without serine treatment (mean ± SD, n = 3 independent experiments). g Generation of CD41a⁺CD42b⁺ MKs from CD34⁺ cells treated with DMSO or the SAHH inhibitor 3DZA with or without serine treatment (mean ± SD, n = 3 independent experiments). h Generation of CD41a⁺CD42b⁺ PLPs from CD34⁺ cells treated with DMSO or the SAHH inhibitor 3DZA with or without serine treatment (mean ± SD, n = 3 independent experiments). i Western blot analysis of H3K9me3, H3K36me3, SETDB2 in cells at day 12 treated with vehicle or serine. Experiment was repeated independently for 3 times. j Venn diagram of downregulated genes and peaks in both ATAC-seq and RNA-seq analyses. k Peaks of the promoter of SVIL, FRYL, and SHANK3 in differentiated cells cultured with vehicle or serine at day 12 as revealed with ATAC-seq. Different regions of primers used for ChIP-qPCR in the promoters are in different colors (purple, gray, and red). l The binding of H3K9me3 to the promoters of SVIL, SHANK3, and FRYL in 293T cells cultured with vehicle or serine by using ChIP-qPCR (means ± SD, n = 3 independent experiments). m Generation of CD41a⁺CD42b⁺ PLPs from CD34⁺ cells with overexpression of SVIL exposed to serine (mean ± SD, n = 3 independent experiments). An unpaired two-sided Student’s t-test was used in c, d, e, f, g, h, l, m. Source data are provided as a Source Data file.