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, 2012). Macromolecules traffic between the nucleus and cytoplasm through these pores that fuse the inner and outer nuclear envelope. Protein complexes known as the nuclear pore complex (NPC) are integrated within the nuclear pores and act as gates that restrict the diffusion of larger biomolecules across  the nuclear envelope. With an approximate mass of 125 MDa, the NPC is one of the largest and most complex assemblages of proteins in the eukaryotic cell and is composed of approximately 30 different nucleoporin (Nup) proteins, with ~500�C1000 individual Nups comprising a single NPC (Reichelt et al., 1990; Cronshaw et al., 2002; Hoelz et al., 2011). The NPC is a dynamic and modular [http://www.selleckchem.com/products/ABT-888.html Veliparib clinical trial] structure with eight-fold rotational symmetry and can be divided into three recognizable ring-like structures surrounding the central channel of the nuclear pore: the cytoplasmic ring, the central spoke ring, and the nuclear ring (which make up the symmetrical portion of NPC) (Frenkiel-Krispin et al., 2010). Attached to the cytoplasmic  ring and nuclear ring are 8 proteinaceous filaments which extend into the cytoplasm and nucleus, respectively, with the nuclear filaments converging to form the nuclear basket (Cautain et al., 2015). These extended structures, together, make up the asymmetric portion of the NPC. Nups are categorized as transmembrane, barrier, or scaffold Nups based upon location within the NPC, amino acid sequence motifs, and structure (Grossman et al., 2012). Transmembrane Nups anchor the NPC to the nuclear envelope pores, barrier Nups facilitate active transport of cargoes, and scaffold Nups  link the transmembrane Nups to the barrier Nups, providing the structural framework of the NPC (Figure ?(Figure11). Figure 1 The nuclear pore complex. The cytoplasmic (dark blue), central spoke (light blue), and nuclear ring (chartreuse) structures constitute the symmetric portion of the nuclear pore complex (NPC) that surrounds the central channel. The asymmetric portion of ... Barrier Nups contain repeated phenylalanine-glycine-rich (FG) sequences that form intrinsically disordered motifs and act as the major impediment to free diffusion through the main channel of the NPC (Cautain et al., 2015). Concomitantly, these FG-Nups provide the only route for active transport of cargo biomolecules between the cytoplasm and nucleus by providing binding sites for nuclear transport receptors, within the NPC, through multiple low-affinity interactions (Ben-Efraim and Gerace, 2001; Ribbeck and G?rlich, 2001). The translocation of complexes through the NPC is energy-independent as GTP hydrolysis is required only as a final step in the transport process (Schwoebel et al., 1998).
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, 2007). Segments were incubated in oxygenated (95% O2; 5% CO2) bicarbonate buffer at 37��C. Tissue  was equilibrated for 15 min, thereafter a 3 mA current pulse was applied across the intestinal wall every 6 s for 30 min. The trans-epithelial potential was measured and recorded by the Acquire and Analyze software, and the change in potential induced by the current pulse was used to calculate trans-epithelial resistance (TER) according to Ohm's Law. Higher values of TER represent lower gut permeability. Immunohistochemical staining Four micro miter-thick cross sections of [http://www.selleckchem.com/JAK.html Selleckchem JAK inhibitor] formalin-fixed, paraffin-embedded ileum and colon tissue were cut, deparaffinized, and subjected to a heat-induced epitope retrieval step. Slides were rinsed in cool running water, washed in Tris-buffered saline (pH7.4) before incubation with primary polyclonal antibody against mouse Muc2 (USCN, USA, dilution 1:100) overnight at 4��C. For detection, rabbit anti-rat secondary antibody was used followed by application of the peroxidase kit (USCN, USA). Alkaline phosphatase was  revealed by Fast Red as chromogen and peroxidase was developed with a highly sensitive diaminobenzidine (DBA) chromogenic substrate for approximately 10 min. Negative controls were performed by omitting the primary antibody. Statistical analysis The relative position and intensity of DNA bands on DGGE profiles were used for principal component analysis (PCA) by SPSS 16.0 statistical package (SPSS Inc. IL, USA). Statistical analyses were performed using a two-tailed  Student t-test, with the assistance from GraphPad Prism Program (version 5.01, GraphPad Software Inc., USA). All values were expressed as means �� standard deviation (SD) and the sample sizes. Significance was accepted at p  *p

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, 2007). Segments were incubated in oxygenated (95% O2; 5% CO2) bicarbonate buffer at 37��C. Tissue was equilibrated for 15 min, thereafter a 3 mA current pulse was applied across the intestinal wall every 6 s for 30 min. The trans-epithelial potential was measured and recorded by the Acquire and Analyze software, and the change in potential induced by the current pulse was used to calculate trans-epithelial resistance (TER) according to Ohm's Law. Higher values of TER represent lower gut permeability. Immunohistochemical staining Four micro miter-thick cross sections of Selleckchem JAK inhibitor formalin-fixed, paraffin-embedded ileum and colon tissue were cut, deparaffinized, and subjected to a heat-induced epitope retrieval step. Slides were rinsed in cool running water, washed in Tris-buffered saline (pH7.4) before incubation with primary polyclonal antibody against mouse Muc2 (USCN, USA, dilution 1:100) overnight at 4��C. For detection, rabbit anti-rat secondary antibody was used followed by application of the peroxidase kit (USCN, USA). Alkaline phosphatase was revealed by Fast Red as chromogen and peroxidase was developed with a highly sensitive diaminobenzidine (DBA) chromogenic substrate for approximately 10 min. Negative controls were performed by omitting the primary antibody. Statistical analysis The relative position and intensity of DNA bands on DGGE profiles were used for principal component analysis (PCA) by SPSS 16.0 statistical package (SPSS Inc. IL, USA). Statistical analyses were performed using a two-tailed Student t-test, with the assistance from GraphPad Prism Program (version 5.01, GraphPad Software Inc., USA). All values were expressed as means �� standard deviation (SD) and the sample sizes. Significance was accepted at p *p

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