Fig. 6: Molecular mechanism of ALKBH5 in regulating human pancreatic lineage specification.

a Flowchart of ALKBH5 downstream target screening. b Venn diagram showing the integrative analysis to identify potential ALKBH5 targets. A5-KO-down: significantly downregulated genes in A5-KO PPs. A5-KO-m⁶A-hyper: genes with significantly higher m⁶A abundance in A5-KO PPs. c GO categories of the positive targets of ALKBH5, p-values were calculated by one-tailed hypergeometric test. d RIP-qPCR showing the m⁶A enrichment increased on indicated mRNA transcripts in A5-KO PPs compared with WT PPs (n = 3 biological replicates). e RIP-qPCR analysis of the indicated mRNA transcripts bound with ALKBH5 in PPs (n = 3 biological replicates). f Violin plots showing expression changes between A5-KO and WT PPs for methylated (with m⁶A) and not-methylated (without m⁶A) transcripts. The upper and lower quartiles and the median are indicated for each group. Dots, the average value of fold changes. p-value was calculated using two-tailed Wilcoxon test. g Cumulative distributions of global transcript half-life changes in WT and A5-KO PPs. h Cumulative distributions of mRNA input changes (A5-KO/WT, log₂FC) of methylated (with m⁶A) and not-methylated (without m⁶A) transcripts. i Cumulative distributions of mRNA lifetime changes (A5-KO/WT, log₂FC) of methylated (with m⁶A) and not-methylated (without m⁶A) transcripts. j Volcano plot of transcripts with significant half-life change in A5-KO PPs compared to WT PPs. Red dots: transcripts with increased half-life; blue dots: transcripts with shortened half-life. k GO categories of genes with shortened half-life, p-values were calculated by one-tailed hypergeometric test. l The decay curves for PDX1, NKX6.1, MNX1, and SOX9 in WT and A5-KO PPs (n = 3 biological replicates). All data are presented as mean ± s.d. Statistical significance calculated using two-tailed Student’s t-test, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Source data are provided as a Source Data file.