Fig. 7: LncBAR facilitates cardiomyocyte proliferation by upregulating BRG1 protein levels. | npj Regenerative Medicine

Fig. 7: LncBAR facilitates cardiomyocyte proliferation by upregulating BRG1 protein levels.

From: Long noncoding RNA LncBAR enhances BRG1 protein to promote cardiomyocyte cell cycle progression and cardiac repair

Fig. 7: LncBAR facilitates cardiomyocyte proliferation by upregulating BRG1 protein levels.The alternative text for this image may have been generated using AI.

a RT-qPCR analysis of Brg1 expression in AR7 hearts. RZ remote zone, BZ border zone, AZ apex zone (n = 3 mice). b RT-qPCR analysis of Brg1 expression (relative to GAPDH) in NMCMs following LncBAR knockdown or overexpression, with shNT or LacZ as controls respectively (n = 3 biological replicates). c Western blot and quantification of BRG1 protein levels in NMCMs treated with or without shLncBAR (n = 3 biological replicates). d Western blot and quantification of BRG1, BAF155 and BAF250a in NMCMs upon LncBAR overexpression, with LacZ as control (n = 3 biological replicates). Western blot analysis (e) and quantification (f) of BRG1 expression in LncBAR-overexpressing NMCMs treated with cycloheximide (CHX, 10 µg/ml) for indicated times (n = 3 biological replicates). Western blot analysis (g) and quantification (h) of BRG1 expression in LncBAR-depleted NMCMs treated with proteasome inhibitor MG132 (10 µM), or lysosomal inhibitors chloroquine (CQ, 10 µM) and NH4Cl (25 mM) for 6 h (n = 3 biological replicates). i RT-qPCR analysis of Brg1 expression in shBrg1-transduced NMCMs, with shNT as control (n = 3 biological replicates). Representative ICC and quantification of cTnT+ Ki67+ (j) and cTnT+ pH3+ (k) cells in NMCMs overexpressing LncBAR, treated with either shBrg1 or shNT control. White arrows indicate proliferating NMCMs (n = 15 fields). Scale bars, 100 µm. l RT-qPCR analysis of cell cycle genes in LncBAR-overexpressing NMCMs treated with shBrg1 or shNT lentiviruses (n = 3 biological replicates). ChIP-seq profiling showed the binding signal for BRG1 at the genomic loci of MCM3 (m) and AURKA (n) promoter regions in reported Nalm-6 cell line31. Western blot (o) and quantification (p, q) of total and phosphorylated PI3K and AKT in NMCMs with LncBAR overexpression and Brg1 knockdown (n = 3 biological replicates). Data are presented as mean ± SEM. Statistical significance was determined by two-tailed unpaired t test, one-way ANOVA, or two-way ANOVA with Tukey’s multiple comparisons test. *P < 0.05, **P < 0.01, ***P < 0.001.

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