Extended Data Fig. 2. Heterogeneity of exon 3 to 6 skipping splicing of Carm1 pre-mRNAs.
a. Percentage of ESS events in mouse two- and four-cell embryos in total (left) or in each blastomere (right) sequenced via PacBio platform. b. The combination charts of bar plots depicting percentage of alternative splicing of Carm1 pre-mRNAs (E3S, E5S, E6S, E56S, E3456S, E4A3SS, E11S, E15S, RI14, and RI15) in mouse late two- and early four-cell embryos and dot plots depicting the relative level of alternative splicing of Carm1 pre-mRNAs among blastomeres in each embryo. Each dot or column respects one blastomere. noAS, non-alternative splicing. L, DNA ladder. Note that the results for exon 3 to 6 skipping splicing are also used in Extended Data Fig. 9b (L2C) and Extended Data Fig. 9c (E4C), and the spatial position information are shown in Extended Data Fig. 9b, c but not here. c, d. The dot plot depicting Carm1 RNA levels based on RNA-seq data from two-cell embryos (n = 42) and four-cell embryos (n = 26) (c), and the qPCR results (d). e. The western blot results showing that the exon-skipping events of exons 3 to 6, but not exon 11 or 15, of Carm1 pre-mRNAs seem to affect the protein level of CARM1. Top panel, illustration of exons of Carm1 pre-mRNA. Bottom left panel, illustration of the vector from which HA-tagged CARM1 and green fluorescent protein (GFP) are expressed simultaneously, in which GFP works as a reference for exogenous transfection. Bottom middle panel, the western blot results. Bottom right panel, the combination charts of bar plots and dot plots showing the relative intensities of HA to GFP (HA/GFP) and GFP to ACTB (GFP/ACTB). EF1α, the promoter of human translation elongation factor 1 alpha. CMV, the promoter of cytomegalovirus. pA, the polyadenylation signal element. kD, kilodalton.