Fig. 6: LncBAR overexpression positively regulated cardiomyocyte survival and proliferation.

a Schematic depiction of experimental design using NMCMs treated with shLncBAR or shNT for RNA-seq (n = 2 biological replicates). b Volcano plot showing differentially expressed genes (DEGs) induced by LncBAR knockdown (shLncBAR) in NMCMs compared to those treated with shNT. DEGs were defined by P < 0.05 and |fold change| ≥ 2 comparing shLncBAR-NMCMs to shNT-NMCMs. c Gene Ontology (GO) analysis of significantly enriched pathways in shLncBAR-treated NMCMs compared with shNT-treated NMCMs. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showing significantly upregulated (d) and downregulated (e) pathways enriched in shLncBAR-NMCMs compared to shNT control cells. f Heatmap showing the expression levels of representative Brg1 downstream factors involved in proliferation in shLncBAR- or shNT-NMCMs (n = 2 biological replicates). g–i Western blot showing the expression of total and phosphorylated form of PI3K and AKT in NMCMs treated with or without LncBAR overexpression. Quantification of indicated protein expression was shown in (h) and (i) (n = 3 biological replicates). Representative ICC images and quantification for cTnT+ pH3+ (j) and cTnT+ Ki67+ (k) NMCMs treated with LncBAR along with small molecule LY294002 (PI3K/AKT pathway inhibitor). Cells infected with lentiviral LacZ and treated with DMSO were used as control. White arrows indicate proliferating NMCMs (n = 15 fields). Scale bars, 100 µm. l RT-qPCR analysis showing expression of genes involved in positive regulation of cell cycle in LncBAR overexpressed NMCMs treated with or without LY294002 inhibitor (n = 3 biological replicates). Data are presented as mean ± SEM. Statistical significance was determined by two-tailed unpaired t test or two-way ANOVA with Tukey’s multiple comparisons test. *P < 0.05, **P < 0.01, ***P < 0.001.