Fig. 1. Both KD and KO of Nr5a2 led to morula arrest.
a TF motifs identified from distal ATAC-seq peaks17,28 in mouse early embryos. Sizes of circles indicate the –LogP value. Expression levels of TFs are color-coded. Epi, E6.5 Epiblast; Ect, E7.5 ectoderm. b Sequence logos of the binding motifs of NR5A2, RARG, NR1H3, and NR2C2 (left), and RNA expression (FPKM) of Nr5a2, Rarg, Nr1h3, and Nr2c2 (right). VE, visceral endoderm. The error bars denote the standard deviations of two biological replicates of RNA-seq. c Bar charts showing the expression of Nr5a2, Rarg, Nr1h3, and Nr2c2 in the negative control (NC) group (injected with negative control siRNA) and siRNA targeting group based on RNA-seq. The error bars denote the standard deviations of two biological replicates of RNA-seq. d Immunofluorescence of NR5A2 (red) and DAPI (blue) in mouse 8C embryos after KD of Nr5a2 (left). Scale bars: 20 μm. Quantification of NR5A2 signal intensity (relative to DAPI) and P values (t-test, two-sided) are shown (right, n = 12–13 embryos). Each dot represents a single blastomere. e Bar plots showing the developmental rates of NC group, Nr5a2 KD group, Rarg KD group, Nr1h3 KD group, and Nr2c2 KD group at the blastocyst stage (E4.5). MO, morula; BL, blastocyst. f Bar charts showing the percentage of WT Nr5a2 (blue) and base-edited Nr5a2 (red) reads from RNA-seq at the 8C stage after injection of Nr5a2 sgRNA only, base-editor (BE) mRNA only, and both sgRNA and BE mRNA. Each bar represents a single embryo. g Immunofluorescence of NR5A2 (red) and DAPI (blue) in mouse 8C embryos after injection of sgRNA only, BE mRNA only, and both sgRNA and BE mRNA (left). Scales bars: 20 μm. Quantification of NR5A2 signal intensity (relative to DAPI) and P values (t-test, two-sided) are shown (right, n = 6–8 embryos). Each dot represents a single blastomere. h Bar plots showing the developmental rates at the blastocyst stage (E4.5) after injection of sgRNA only, BE mRNA only, and both sgRNA and BE mRNA.