Fig. 2: Echs1 is the key regulator of histone Kcr in early mouse embryos. | Nature Communications

Fig. 2: Echs1 is the key regulator of histone Kcr in early mouse embryos.

From: Echs1-mediated histone crotonylation facilitates zygotic genome activation and expression of repetitive elements in early mammalian embryos

Fig. 2

A Model diagram of key factors controlling histone crotonylation modification. Crotonyl-CoA and β-hydroxybutyryl-CoA can be interconverted by Echs1. In addition, crotonyl-CoA can be generated from crotonic acid catalyzed by Acss2 or from Butyryl-CoA catalyzed by Acox3, and ultimately converted unidirectionally to β-hydroxybutyryl-CoA catalyzed by Cdyl. B Experimental design schematic for Echs1 knockdown in oocytes, IVM, IVF, and preimplantation development of mouse embryos. Generally, siRNA was injected into GV oocytes for IVM, then MII oocytes were obtained for IVF, the zygotic and preimplantation embryos were obtained through culturing in KSOM medium. C Dotplot indicated expression changes of known histone Kcr writers, readers, and regulators in mouse oocytes and preimplantation embryos (log2FPKM). The transcriptome data was derived from GSE165782. D Immunofluorescence staining and MFI of ECHS1 protein in early 2-cell embryos of siCtrl (control siRNA was injected) and siEchs1 (siRNA against Echs1 was injected) groups (n = 16). Scale bar: 20 μm. E Examination and quantification analysis of ECHS1, H3K9cr, and total histone H3 protein levels in siCtrl and siEchs1 early 2-cell embryos by immunoblotting. The blots were derived from the same experiment and processed in parallel. Total histone H3 level was set as an internal reference (n = 300). F, G Immunofluorescence staining and MFI of H3K9cr (F) (n = 16) and H3K4cr (G) (n = 27) in siCtrl and siEchs1 embryos. Scale bar: 20 μm. H Representative developmental progression images of early embryos. Different stage embryo percentages were shown to average on the right (n  =  3 independent replicates). Scale bar: 100 mm. I, J Confocal images showed newly synthesized DNA by EU staining (I) (n = 40) and newly synthesized RNA by EdU staining (J) in siCtrl and siEchs1 embryos (n  =  3 independent replicates), scale bar: 20 μm, MFI and the percentages of positive EdU embryos was shown on the right of each image. In (D), (E), (F), (G), (I) and (J), data were presented as Mean ± SD of three independent replicates and statistical evaluation was performed by two-tailed unpaired Student’s t-test. Source data are provided as a Source Data file.

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