Fig. 4

MIDEAS-AS1 activates NCALD expression via recruiting the MATR3 complex to the NCALD promoter. A Heat map of differentially expressed gene based on RNA-seq analysis between MIDEAS-AS1-overexpression cells and control cells. B There were 471genes differentially expressed between MIDEAS-AS1-overexpression and control (252 up and 219 down) cells (|log2(Fold change) |> 1). C Top 20 up-regulated genes were showed. D Venn-diagrams showing intersect between down-regulated genes in TCGA data (1648 genes) and up-regulated DEGs after MIDEAS-AS1-overexpressing data (252 genes). E–F qRT-PCR and Western blot analysis of NCALD expression in MDA-MB-231cells and MDA-MB-468 cells with knockdown or overexpression MIDEAS-AS1 or MATR3. G qRT-PCR analysis of the expression of NCALD in MDA-MB-231 and MDA-MB-468 cells co-transfected with MIDEAS-AS1 overexpression vector or empty vector together with si-MATR3 or si-NC. H Western blot analysis of the expression of NCALD in MDA-MB-231 and MDA-MB-468 cells co-transfected with MIDEAS-AS1 overexpression vector or empty vector together with si-MATR3 or si-NC. I Luciferase reporter assays validated the binding of MIDEAS-AS1 and MATR3 with NCALD. J Luciferase assays in HK293T cells was determined by co-transfected with MIDEAS-AS1-overexpressing vector and si-MATR3. K-L Relative luciferase activity of full-length promoter and the other five truncated promoter regions of NCALD in HK293T cells by transfected with MIDEAS-AS1-overexpressing vector. M ChIP-qPCR experiments on ten different NCLAD promoter primer using anti-Flag antibody in MDA-MB-231 and MDA-MB-468 cells transfected with MIDEAS-AS1 overexpression plasmid and N-terminal FLAG-tagged MATR3 plasmid. Data were shown as mean ± SD. (*p < 0.05, ** p < 0.01, *** p < 0.001)