RNA-seq and Cut & Tag analyses of mouse 2-cell embryos in which lamin B1 dissociation from the nuclear envelope is inhibited

This lamin B1 dissociation is mediated by autophagy and can be inhibited by interfering with lamin B1’s binding to LC3, a factor responsible for early autophagosome formation. In order to block interaction between lamin B1 and LC3, we injected mRNA harboring the LC3-binding domain of lamin B1 to mouse embryos and examined transcriptomes of the injected embryos by RNA sequencing (RNA-seq) analysis.

In addition, embryos expressed with full-length lamin B1 mRNA harboring two different types of mutations (flLB1(F454A/L457A) and flLB1(5A)) were analyzed by RNA-seq, together with non-injection control (Non2). Another injection control (flLB1(F454A/L457A)ΔC) was also performed, together with a non-injection control (Non3). Furthermore, the effect of nuclear stiffening was examined by expressing the molecular tether-anchor system (tWT), together with a mutant carrying LaG165A (tMut) and a non-injection control (Non4).