Fig. 3: bnRNA-seq analysis comparing newly fused myonuclei acquired during developmental and adult growth.

a Schematic diagram of the bulk nuclei RNA-seq of GFP⁺ and GFP⁻ myonuclei from Pax7^rtTA; TRE^H2B-GFP mice during postnatal development. TA and gastrocnemius muscles at P28 were used after 10 days of doxycycline treatment. b Principal component analysis of the top 500 variable genes from bnRNA-seq of newly fused myonuclei (GFP⁺) and pre-existing myonuclei (GFP⁻). c Volcano plot depicting DEGs between newly fused myonuclei (GFP⁺) and pre-existing myonuclei (GFP⁻). d Gene Ontology (GO) analysis of the DEGs from (c), showing significantly changed biological processes with a false discovery rate (FDR) < 0.01. Gene lists used for GO analysis are shown in Supplementary Data 2. e Schematic diagram for bnRNA-seq of GFP⁺ and GFP⁻ myonuclei from Pax7^rtTA; TRE^H2B-GFP mice during adult muscle overload. f Principal component analysis of bnRNA-seq data from newly fused myonuclei (GFP⁺) and existing myonuclei (GFP⁻) isolated from plantaris muscle 1 week after muscle overload, compared with sham myonuclei (Pax7^rtTA; TRE^H2B-GFP mice treated with Dox as in (e)) and developmental myonuclei datasets from (b). g Volcano plots reveal up- and down-regulated DEGs in newly fused myonuclei (GFP⁺) or pre-existing myonuclei (GFP⁻) compared with sham myonuclei. h Volcano plot reveals up- and down-regulated DEGs from comparison of GFP⁺ and GFP⁻ myonuclei.