Fig. 8: The m6A reader IGF2BP2 is involved in the maturation defects caused by Alkbh5 knockout.
a The density plot showing the distance between binding sites of m6A readers (IGF2BP2, IGF2BP3, YTHDF2 and YTHDC1) detected by RIP-seq and eCLIP-seq in mouse and human ESCs and m6A peaks of loss-GMD detected by m6A-seq in our dataset. b–e RIP-qPCR results showing the relative enrichments of the indicated m6A readers among the targeted genes in oocytes. The data validated by qPCR are present as mean ± SD and are normalized to IgG samples. For P values in c P = 0.000092, 0.000239 and 0.000020; in e P = 0.000302, 0.000439 and 0.000107 (from left to right). f–i RIP-qPCR results showing the relative enrichments of IGF2BP2 among the targeted genes in WT and Alkbh5−/− MII oocytes. The data are presented as mean ± SD and are normalized to IgG samples. j Bubble plot showing the expression fold changes of Igf2bp1, Igf2bp2 and Igf2bp3 during oocyte maturation upon Alkbh5 depletion detected by RNA-seq. Genes differential expression analysis was performed by DESeq2 software between two different groups. k RT-qPCR results showing the knockdown efficiency of Igf2bp2 RNA interference in WT and Alkbh5−/− oocytes respectively. The bar is mean ± SD resulting from three independent experiments. WT, P = 0.000034. l The rates of GVBD and PB1 emission in WT and Alkbh5−/− oocytes microinjected with control and Igf2bp2 siRNA. Numbers of oocytes examined are indicated. Data are presented as mean values ± SD. GVBD, P = 8.78e−11; PBE, P = 0.000068. m Representative confocal images of spindle assembly and chromosome alignment in oocytes microinjected with control and Igf2bp2 siRNA, respectively. Spindle is labeled in green by α-Tubulin, while DNA is marked in blue by DAPI. Phalloidin (red) highlights the margin of oocyte. n–p RT-qPCR results showing the relative mRNA levels of the indicated genes in MII oocytes microinjected with control and Igf2bp2 siRNA. The bar is mean ± SD resulting from three independent experiments. For P values in n P = 0.000043; in p P = 0.00002. Data in b-i represent results from three biologically independent experiments. P values in b-i, k-l and n-p were calculated by two-tailed Student’s t test. Source data are provided as a Source Data file.
