Fig 2: SsD inhibits patient primary cells. (A) CCK-8 assays in the cells derived from two patients treated with SsD for 48 h. (B) Colony-forming assays of patient cells treated with SsD. Graphs showing the colony number from 3 independent experiments. Data are the mean ± SD; *P < 0.05, **P < 0.01. (C) Effect of SsD on cell-cycle arrest was determined using FACS based on PI staining in patient cells after 48 h of treatment. (D) After 48 h of SsD treatment, m6A abundance in the RNA samples was determined in primary patient cells using a dot blot assay. (E) TSQ traces of FTO demethylation of m6A in ssRNA in the absence and presence (1 µM SsD for 48 h) of SsD in patient cells. (F) qPCR expression analysis of the indicated genes in patients' SsD-treated cells. Data represent three independent experiments. (G) Western blot analysis of the indicated genes in patients' SsD-treated cells. (H) Leukemia-bearing mice were developed by intravenous injection of patient cells into 4-week old NSG mice. (I) WBC count of leukemia-bearing mice (n = 3) is shown. (J) Images show representative external views of the spleen with the histological analysis of H&E-stained spleen sections (Scale bars, 1 cm, middle), Wright-Giemsa-stained bone marrow (BM) cells (left), and external views of the lung with H&E-stained sections (right) from leukemia-bearing mice. (K-M) The quantification of blast cells (K), spleen weight (L), and tumor growing nodules on lung (M) are shown. (N) The survival curve of leukemia-bearing mice calculated by Kaplan-Meier survival analysis (n = 5) is shown.

Fig 3: METTL3 promotes GC metastasis in vivo. (A) LV-NC, LV-METTL3 and LV-METTL3+shDEK cells were injected into nude mice via tail vein, and the growth of lung metastases was monitored by bioluminescence imaging 28 days later (n = 3). (B) Statistical analysis of luminescence values of nude mice bioluminescence imaging to monitor lung metastases (**: p < 0.01). (C) Representative image of nude mouse lung tissue for HE staining to evaluate lung metastases.

Fig 4: The weakened binding affinity of METTL3 in POCD. The protein levels of METTL3, METTL14, WTAP, ALKBH5, FTO, YTHDF1, IGF2BP1 and HNRNPA2B1 in the hippocampuses of POCD mice. (A) The binding affinity of METTL3 on BDNF, SOX2, and SYN1, (B) CCND2, STAR, and TIGAR, (C), as well as BACE1 and IL17A (D) in hippocampal neurons of POCD mice shown by RIP-qPCR. The binding affinity of FTO, (E) ALKBH5 (F) and IGF2BP1 (G) on BDNF, SOX2, and SYN1 in hippocampal neurons of POCD mice shown by RIP-qPCR. Data are presented as the mean ± standard error of the mean of three individual experiments; p < 0.05 vs. control group.

Fig 5: Differential m6A RNA methylation at the 5′UTR in POCD. The genes with differential m6A RNA methylation at the 5′UTR in POCD mice depicted by a volcano plot. (A) The GO analysis of these genes is depicted by a bubble chart. (B) The m6A enrichment at the 5′UTR of SOX2, SYN1, BDNF, BACE1, and IL17A in POCD mice depicted by IGV. (C) The mRNA levels of SOX2, SYN1, BDNF, BACE1, and IL17A in POCD. (D) The m6A enrichment at the 3′UTR of CCND2, STAR, and TIGAR in POCD shown by IGV. (E) The mRNA levels of CCND2, STAR, and TIGAR in POCD. (F) Data are presented as the mean ± standard error of the mean of three individual experiments; p < 0.05 vs. control group.