Control (a), dilated (b) and ischaemic tissue (c). The labeling is INCB-039110 biological activity increased in ischaemics. No 23388095 labely of NDC1 was observed in other nuclear structure. Scale bar = 50 nm. (B) Western blot analysis of NDC1, Nup160 and Nup93 in nuclear (Nu) and cytosolic (Cy) fraction in controls and HF patients (ICM and DCM). doi:10.1371/journal.pone.0048957.gand Nup93 in HF patients when compared to the control group. These results, together with the data observed in other studies, suggest a relationship between changes in nucleoporin levels and the alterations of the nucleocytoplasmic transport previously reported. It has been shown that overexpression of both Nup160 and Nup153 causes inhibition of mRNA export, observing nuclear accumulation of RNA poly(A)+; Triptorelin however, the import and export of proteins is not affected, so it seems that 23115181 no nucleocytoplasmic transport reduction occurs, nor does occlusion or disassembly of the NPC [23?4]. The results obtained by Vasu et al. [24], showed that Nup160 could interact directly with specific factors or receptors involved in mRNA export and/or indirectly immobilize Nup153 in the pore, which has a region representing the docking site for mRNA molecules [23]. Thus, our results are consistent with these previous data. Moreover, despite the significant increases in protein levels of Nup160 and Nup93 in HF patients compared with the control group, the immunofluorescence distribution patterns were similar in all the groups studied, these proteins being located in the nuclear envelope. Nup160 has an important structural role, participating in the initial stages of NPC assembly [26], and it is also involved in the transport of mRNA [24]. All these processes take place in the nuclear envelope, which could determine the subcellular localization of Nup160, especially in patients with HF where the nucleocytoplasmic transport is altered. Nup93 also has an important structural role since its depletion results in deformed pores, and together with the depletion of NDC1 causes a global disruption NPC [27]. Also, it has been seen that Nup93 is associated with important regions of the chromatin involved in transcriptional regulation [28]. As with Nup160, these are important functions that take place in the nuclear envelope and may determine the subcellular localization of Nup93. Stability in the subcellular localization of Nup93 has been confirmed in independent studies [28?9]. In addition, we have observed two distribution patterns in NDC1 protein. NDC1 is observed on the nuclear surface and inside the nucleus. Previous studies show a similar pattern in the control group of NDC1 in the nuclear surface [27]. NDC1 is an evolutionarily conserved transmembrane protein in most eukaryotes [27], necessary for the proper assembly of the NPC and its absence causes severe defects in the assembly and NPC biogenesis [30?1]. As described in HeLa cell cultures, we observed that NDC1 is located at the nuclear envelope in the CNT group [27]. However, we also observed a distribution within the nucleus in the ICM and DCM groups. It has been observed that in yeast increased Nup53p (hNup35) nucleoporin locates in the core, causing the accumulation of NDC1 in lamellar membranes inside the nucleus. In this sense, we believe that this protein could be related to Nup160, also a core protein increased in the HF patients group, sequestering NDC1 into the nucleus. Furthermore, the assembly of the components of the NPC is a complex and little known process tha.Control (a), dilated (b) and ischaemic tissue (c). The labeling is increased in ischaemics. No 23388095 labely of NDC1 was observed in other nuclear structure. Scale bar = 50 nm. (B) Western blot analysis of NDC1, Nup160 and Nup93 in nuclear (Nu) and cytosolic (Cy) fraction in controls and HF patients (ICM and DCM). doi:10.1371/journal.pone.0048957.gand Nup93 in HF patients when compared to the control group. These results, together with the data observed in other studies, suggest a relationship between changes in nucleoporin levels and the alterations of the nucleocytoplasmic transport previously reported. It has been shown that overexpression of both Nup160 and Nup153 causes inhibition of mRNA export, observing nuclear accumulation of RNA poly(A)+; however, the import and export of proteins is not affected, so it seems that 23115181 no nucleocytoplasmic transport reduction occurs, nor does occlusion or disassembly of the NPC [23?4]. The results obtained by Vasu et al. [24], showed that Nup160 could interact directly with specific factors or receptors involved in mRNA export and/or indirectly immobilize Nup153 in the pore, which has a region representing the docking site for mRNA molecules [23]. Thus, our results are consistent with these previous data. Moreover, despite the significant increases in protein levels of Nup160 and Nup93 in HF patients compared with the control group, the immunofluorescence distribution patterns were similar in all the groups studied, these proteins being located in the nuclear envelope. Nup160 has an important structural role, participating in the initial stages of NPC assembly [26], and it is also involved in the transport of mRNA [24]. All these processes take place in the nuclear envelope, which could determine the subcellular localization of Nup160, especially in patients with HF where the nucleocytoplasmic transport is altered. Nup93 also has an important structural role since its depletion results in deformed pores, and together with the depletion of NDC1 causes a global disruption NPC [27]. Also, it has been seen that Nup93 is associated with important regions of the chromatin involved in transcriptional regulation [28]. As with Nup160, these are important functions that take place in the nuclear envelope and may determine the subcellular localization of Nup93. Stability in the subcellular localization of Nup93 has been confirmed in independent studies [28?9]. In addition, we have observed two distribution patterns in NDC1 protein. NDC1 is observed on the nuclear surface and inside the nucleus. Previous studies show a similar pattern in the control group of NDC1 in the nuclear surface [27]. NDC1 is an evolutionarily conserved transmembrane protein in most eukaryotes [27], necessary for the proper assembly of the NPC and its absence causes severe defects in the assembly and NPC biogenesis [30?1]. As described in HeLa cell cultures, we observed that NDC1 is located at the nuclear envelope in the CNT group [27]. However, we also observed a distribution within the nucleus in the ICM and DCM groups. It has been observed that in yeast increased Nup53p (hNup35) nucleoporin locates in the core, causing the accumulation of NDC1 in lamellar membranes inside the nucleus. In this sense, we believe that this protein could be related to Nup160, also a core protein increased in the HF patients group, sequestering NDC1 into the nucleus. Furthermore, the assembly of the components of the NPC is a complex and little known process tha.
