Quantitative proteomics analysis reveals the pathogenesis of obstructed defecation syndrome caused by abnormal expression of dystrophin

Figure 4 Overexpression of nucleoporin protein 153 and L-type voltage-gated calcium channel is associated with downregulation of dystrophin expression in human intestinal smooth muscle cells. A: Real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression of dystrophin (DMD) in human intestinal smooth muscle cells transfected with negative control small interfering RNA (siR-NC) or siR-dystrophin (siR-DMD); B: RT-qPCR was used to detect the expression of nucleoporin protein 153 (NUP153) in human intestinal smooth muscle cells transfected with siR-NC or siR-DMD; C: RT-qPCR was used to detect the expression of L-type voltage-gated calcium channel (Ca_v1.2) in human intestinal smooth muscle cells transfected with siR-NC or siR-DMD; D: Western blot analysis of the levels of DMD, NUP153, and Ca_v1.2 in human intestinal smooth muscle cells transfected with siR-NC or siR-DMD; E: Quantitative analysis of Western blot greyscale value; F: Representative immunohistochemistry staining for NUP153 and Ca_v1.2 in rectal specimens from two groups of 50 cases. Scale bar = 50 μm; G: Quantitative analysis of the immunohistochemistry expression of NUP153 and Ca_v1.2 (integrated optical density of NUP153 and Ca_v1.2) in rectum samples of the obstructed defecation syndrome (ODS) group compared with the control group; H: Representative immunofluorescence staining for NUP153 in rectal specimens from two groups of 50 cases. Scale bar = 20 μm. Nuclei were counterstained with DAPI; I: Quantitative analysis of mean fluorescence intensity of NUP153 relative to the ODS and control groups; J: Representative immunofluorescence staining for Ca_v1.2 in rectal specimens from two groups of 50 cases. Scale bar = 20 μm. Nuclei were counterstained with DAPI; K: Quantitative analysis of mean fluorescence intensity of Ca_v1.2 relative to the ODS and control groups. ^aP < 0.05, ^cP < 0.001, ^dP < 0.0001, n = 3. siR-NC: Negative control small interfering RNA; siR-DMD: Dystrophin small interfering RNA; DMD: Dystrophin; ODS: Obstructed defecation syndrome; NUP153: Nucleoporin protein 153; Ca_v1.2: L-type voltage-gated calcium channel.