Supplementary Figure 7: Bach TFs may contribute to the development of MDS phenotype.

a, The erythroid differentiation of human cord blood CD34⁺ cells with Ctlr-shRNA or shRNA targeting BACH1 or BACH2 were analyzed by flow cytometry. CD71⁺GlyA⁺ cells were considered as differentiated erythroid cells. Three different shRNAs targeting BACH1 or BACH2 were used as shown in the dot plot (Ctlr n = 1 vector, Bach1 n = 3 vectors, Bach2 n = 3 vectors, Data obtained from three experiments were shown). b, RNA-based next-generation sequeincing analysis of indicated genes in CD34⁺ cells from healthy control (HC, n = 3) or from MDS patients (MDS, n = 100). Results were shown in violin plot. c, Scatter plot of Bach2 and Rag2 expressions in RNA-based next-generation sequeincing data. The correlation coefficient r = 0.73. d, Enrichments of the Bach1 and Bach2 on Ccl2. e, Results of the serum cytokines (IL-6 and CCL2) analysis (wild-type, n = 6 mice; Bach1^—/—, n = 6 mice; Bach2^—/—, n = 5 mice; each dot represents individual mice. Analysis was done using the samples obtained from two experiments). f, GSEA results of indicated gene set using RNA-based next-generation sequeincing data of MDS patients were shown. MDS patients were divided into two groups depending on their BACH2 expression levels (BACH2_high; higher than median expression levels of BACH2, Bach2 _low; lower than median expression levels of BACH2). ES: enrichment score, P-val: nominal p-value as implemented in GSEA. g,h, Subtype analysis of BACH2 expression levels in MDS. Subtype analysis of the data shown in Figure 8b (g). Subtype analysis of the data shown in Supplementary figure 7b (h). Violin plot and Box-and-whisker plot show median, interquartile range, minimum and maximum values (b,e,g,h). Statistical analysis was done with two-tailed Student’s t test or Welch’s t test (b) and two-sided Jonckheere-Terpstra test (g). No adjustments were made for multiple comparisons.