Research Paper Volume 11, Issue 20 pp 8792—8809

Long non-coding RNA MALAT1 promotes cardiac remodeling in hypertensive rats by inhibiting the transcription of MyoD

Figure 4. LncRNA MALAT1 over-expression suppresses the transcription of MyoD. (A) the expression of MyoD in myocardial tissues and thoracic aortic vascular tissues of SD rats and SHRs detected by Western blot analysis; (B) the expression of MyoD in myocardial tissues and thoracic aortic vascular tissues of SHRs after over-expression or silencing of lncRNA MALAT1; (C) cellular location of lncRNA MALAT1 determined using FISH (× 400); (D) enrichment of lncRNA MALAT1, U1, and GAPDH in nuclear or cytoplasmic fractions of ASMCs; (A) luciferase activity of Myogenin-Luc and MCK-Luc in VMSCs in response to over-expression or silencing of lncRNA MALAT1; (G) the expression of MyoD and MyoG in VSMCs of SHRs after over-expression or silencing of lncRNA MALAT1, as measured by Western blot analysis; (H) RIP assay of nuclear extracts from the VSMCs immunoprecipitated by IgG or an antibody against Suv39h1 and RT-qPCR measurement of the retrieved RNAs; (I) RNA pull-down assay of in vitro-transcribed biotinylated full-length lncRNA MALAT1 transcripts and the binding proteins as well as Western blot analysis of the indicated proteins; (JL) ChIP-PCR analysis of Suv39h1, H3K9me3 and MyoD enrichment on the promoter or enhancer of Myogenin, MCK loci in VSMCs. The enrichment fold was calculated as a fraction of DNA present in the input samples. *, p < 0.05 vs. the SD rats or cytoplasmic RNA or LV-CON-vector group or LV-CON-shRNA group or IgG group; #, p < 0.05 vs. the MyoD or Suv39h1 or H3K9me3; measurement data were expressed by means ± standard deviation and analyzed by unpaired t-test; n = 3.