Fig. 7: YTHDF2 knock-down rescues and dm-αKG promotes human pancreatic differentiation. | Nature Communications

Fig. 7: YTHDF2 knock-down rescues and dm-αKG promotes human pancreatic differentiation.

From: N6-methyladenosine modification-mediated mRNA metabolism is essential for human pancreatic lineage specification and islet organogenesis

Fig. 7

a RIP-qPCR analysis of the indicated mRNA transcripts bound with YTHDF2 in PPs (n = 3 biological replicates). b A schematic representation of the generation of PPs from hPSCs. WT + shNC, A5-KO + shNC, A5-KO + shDF2-1, and A5-KO + hDF2-2 hPSC lines were used. c RT-qPCR analysis of the expression of YTHDF2 (n = 3 biological replicates). d Representative flow cytometry plots and the percentage of PDX1+NKX6.1+ cells in WT + shNC, A5-KO + shNC, and A5-KO + shDF2 PPs (n = 3 biological replicates). e RT-qPCR analysis of PDX1 and NKX6.1 expression (n = 3 biological replicates). f Immunofluorescent staining of PPs for PDX1, NKX6.1, and nuclei. Images are representative of three independent replicates. Scale bar, 100 μm. g RT-qPCR analysis of PDX1, NKX6.1, SOX9, and MNX1 expression (n = 3 biological replicates). h RIP-qPCR showing the m6A enrichment decreased on indicated mRNA transcripts in PP populations treated with dm-αKG (n = 3 biological replicates). i A schematic summary of functions and molecular mechanisms of ALKBH5 during human pancreatic specification and islet organogenesis. All data are presented as mean ± s.d. Statistical significance calculated using two-tailed Student’s t-test, ns p > 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Source data are provided as a Source Data file.

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