Fig. 4 | Nature Communications

Fig. 4

From: miR-103 promotes endothelial maladaptation by targeting lncWDR59

Fig. 4

LncWDR59 interaction with Notch1-inhibitor Numb. a Nuclear and cytoplasmic lncWDR59 levels in MAoECs treated with miR-103 or control-LNA inhibitors for 24 h. B2m was used as quality marker of nucleus and cytoplasmic RNA isolation (n = 3 per group), expressed as percentage of total b2m RNA expression. Gapdh was used for relative quantification. b LncWDR59 expression in MAoECs before (input) and after NICD-immunoprecipitation (IP) analyzed by qPCR and loaded on 2% agarose gel (lncWDR59 amplicon of 92 bp). Efficiency of NICD-IP was tested by western blot (n = 3 per group). IgGs were used as IP-negative controls. c Heat-maps representing the interaction propensity analysis between lncWDR59 and Numb. The y- and x-axes represent the index of the protein and lncWDR59 sequences, respectively. The colors indicate the interaction score of individual nucleotide and amino acid pairs (rank ± 4). The interaction strength with respect to a training set is represented by the interaction score and the discriminative power values. d, e LncWDR59 expression in MAoECs treated with miR-103 or control-LNA inhibitors for 24 h analyzed by qPCR, following Numb-IP. Amplification products were loaded on 2% agarose gel (lncWDR59 amplicon of 92 bp), together with PCR products from lncWDR59 expression analyzed before IP (d). Numb-IP efficiency was evaluated by western blot (d) and NICD binding to Numb quantified (n = 3 per group) (e). IgGs were used as IP-negative controls. FC fold change of control-LNA inhibitors of the corresponded subcellular fraction, IP immunoprecipitate, NICD Notch intracellular domain, aa amino acid, nt nucleotides. *P < 0.05 by Student’s t-test

Back to article page