Fig. 1: Elevated m⁶A and METTL14 level predicted unfavorable prognosis in MDS.

A m⁶A colorimetric quantification of the global m⁶A level in bone marrow mononuclear cells (BM-MNCs) of 29 MDS patients in our center (A–C) showing higher global m⁶A level in MDS with BM ≥ 5% compared with MDS with BM < 5% and healthy donors. B Higher global m⁶A level in MDS with higher-risk (IPSS-R > 3.5) compared with MDS with lower-risk (IPSS-R ≤ 3.5) and healthy donors. C Kaplan–Meier survival analysis showing that MDS patients with high m⁶A level had shorter OS than MDS patients with low m⁶A level. D Gene expression and survival analyses being conducted using the publicly available MDS database (GSE58831) (D–F): the left plot showing the expression profile of 23 m⁶A regulators in bone marrow CD34⁺ cells of MDS patients stratified by the percentages of bone marrow blasts (P^# values based on the Kruskal-Wallis’ test); the right plot showing univariate Cox analysis (P^$ values) of the associations of the expressions of m⁶A regulators with OS in MDS patients. E Multivariate Cox regression showing the independent prognostic significance of m⁶A regulators which got p < 0.1 in univariate analysis. F Venn diagram showing that METTL14 was the only regulator whose expression level was not only related with the percentages of bone marrow blasts but also independently associated with prognosis. G RNA-seq analysis of METTL14 expression in bone marrow CD34⁺ cells obtained from 16 MDS patients and 10 healthy donors in our center showing higher expression of METTL14 in MDS with BM ≥ 5% compared with MDS with BM < 5% or healthy donors. H q-PCR analysis of METTL14 mRNA expression in BM-MNCs obtained from 221 MDS patients in our center (H, I, L, M) showing elevated METTL14 expression in MDS patients with BM ≥ 5% compared to those with BM < 5% or healthy donors, while the expression levels of METTL14 in MDS patients with BM ≥ 5% being similar as those observed in the AML cell lines. I Higher METTL14 expression in MDS with higher-risk (IPSS-R > 3.5) category compared with MDS with lower-risk (IPSS-R ≤ 3.5) category or healthy donors. J Western blot showing METTL14 protein was highly expressed in MDS patients with BM ≥ 5%. K Western blot showing METTL14 protein was highly expressed in MDS cell line MDS-L. L Kaplan–Meier survival analysis showing that MDS patients with high METTL14 level had shorter OS than MDS patients with low METTL14 level. M Kaplan–Meier survival analysis showing that MDS patients with high METTL14 level had shorter LFS than MDS patients with low METTL14 level. The differences of gene expressions of MDS patients were compared using Mann-Whitney’s test or Kruskal-Wallis’s test. Kaplan–Meier survival analysis was performed using log-rank test. Error bars denoted median ± 95% CI. N.S No significance.