Fig. 2. Proteomic and bioinformatic analysis of LGR5 knockdown in SH-SY5Y cells. (A) Coomassie brilliant blue R-350 stained gels after 2D-PAGE. Protein spots in SH-SY5Y cells after treatment with control siRNA or LGR5 siRNA. Common landmarks of protein spots in the SH-SY5Y cells after treatment with control siRNA and LGR5 siRNA are indicated. (B) Identification of protein spots. Protein spots exhibiting a change greater than twofold in the expression in SH-SY5Y cells treated with LGR5 siRNA compared to control cells. (C) Detailed information of the 12 selected protein spots. (D) Proteins with increased and decreased expression after LGR5 inhibition compared to control-treated cells. (E) Identification of different expression levels of hnRNPA2B1 and hnRNPH3 in SH-SY5Y cells after treatment with LGR5 siRNA or control siRNA by western blotting. (F) LGR5-associated protein-protein interaction network. An LGR5-associated protein-protein interaction network was constructed using a bioinformatics approach. Circles indicate LGR5 and the 12 proteins found to be associated with LGR5 in neuroblastoma cells by a proteomics approach. The thickness of the line between any two proteins represents the degree of confidence in the interaction between the two proteins, with thicker lines indicating higher confidence (left upper). The types of interactions between proteins are represented by action types and effects (right lower).
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