Fig. 5

LINCM expression validated by single molecule RNA FISH. a Single nuclear RNA-seq identifies 141 novel lincRNAs in nuclei of CMs (LINCMs) that are not in current public databases (ENSEMBL and NONCODE) nor published cardiac transcriptome data sets. b Single nuclear RNA-seq identifies LINCMs that are not detectable in matched left ventricle bulk tissue RNA-seq, explained by the dilution of reads in cytoplasmic mRNA pool. c Active H3K27Ac enhancer chromatin regions proximal to LINCMs are enriched in MEF2 transcription factor binding motif and functionally annotated by GREAT analysis to have cardiac expression and phenotypes. d Sites of active transcription demonstrated by co-localization of exonic and intronic probes (asterisk) in nucleus. Scale bar represents 5 μm. e–m Single molecule RNA FISH validates the expression of LINCMs in isolated adult mouse CMs. n–q Positive controls for highly abundant core genes Tpm1, Tnnt2, Myl2, and Malat1. r, s Negative controls with no-probe control (NPC) (r) and sense probe (s) to confirm signal specificity. Scale bar represents 10 μm. t, u Gas5 is upregulated in TAC CM and co-localizes with perinuclear Nppa transcripts. v, w Sghrt is upregulated and localizes to the cytoplasm of TAC CM. x, y LINCM5 is downregulated in TAC CM. Scale bar represents 10 μm