Fig. 2: RBPs interact with GRB10 pre-mRNA. | Cell Death & Disease

Fig. 2: RBPs interact with GRB10 pre-mRNA.

From: The circular RNA circ-GRB10 participates in the molecular circuitry inhibiting human intervertebral disc degeneration

Fig. 2

a A schematic drawing of four types of circ-GRB10 overexpressing vectors (#1 to #4). The genomic region for circ-GBR10 (green bars) with its wild-type flanking introns (black lines) was inserted into the pcDNA3.1 expression vector (#1). 2RC and 6RC are indicated by red bars. A series of deletions are indicated by black crosses (#2 to #4). b qRT-PCR showed the expression of circ-GBR10 after transfection with the four types of circ-GRB10 overexpressing vectors (#1 to #4). Three independent repeats were performed in each experiment. ***p < 0.001. c Schematic diagram of RNAs corresponding to different fragments of GRB10 pre-mRNA (P1, P2) produced by in vitro transcription in the presence of biotin for RNA pulldown experiments. d Silver stain acrylamide gel of total nuclear proteins before (Input) and after pulldown with biotin-labeled RNA probe (P1, P2). M, molecular weight marker (kDa). e Proteins identified from mass spectrometry were integrated to STRING database and constructed Protein-protein interaction (PPI) network. A densely connected module which contains 27 proteins, including FUS, was screened from the PPI network, and these proteins were participate in biological process of mRNA splicing, via spliceosome. f Pathways enrichment analysis of proteins in PPI network. g RBPs which can potentially bind circ-GRB10 pre-mRNA.

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