We have on LGMD proteins or open new directions for specific areas of research. For example, the interaction between DYSF, the LGMD2B protein, and SNAPIN, a component of the SNARE complex that is required for vesicle fusion [26] is appealing with regard to the role ofBlandin et al. Skeletal Muscle 2013, 3:3 http://www.skeletalmusclejournal.com/content/3/1/Page 9 ofTable 3 Most frequent domains in the LGMD-centered dataset and representativeness in the human proteomeDomains Immunoglobulin (IPR003599; IPR003598; IPR007110) Zinc finger, C2H2-type (IPR007087) Fibronectin type III (IPR003961) Ankyrin repeat (IPR002110) Nebulin 35 residue motif (IPR000900) Midostaurin in the total of SID 22.5 (n=141) 12.8 (n=80) 9.63 (n=60) 5.3 (n=33) 4.5 (n=28) in the human proteome (according to M ler et al., Genome Res 2002) 4.2 (n=1214; r=2) 17.6 (n=5092; r=1) 2.9 (n=842; r=5) 0.9 (n=278; r=14) n.i.(n.i.= not indicated; n= number of occurrences; r= rank in the human proteome).DYSF in membrane repair [27]. The positive interaction between kinectin (KTN1), a kinesin anchoring protein that modulates the endoplasmic reticulum interaction with the microtubule and that is known to interact with Rho GTPases [28], and ABI1, an adaptor protein involved in the transduction of signals from Ras to Rac and the regulation of actin polymerization [29], suggests a role for this interaction in the regulation of the muscle cytoskeletal remodeling. We then investigated intracellular co-localization on normal human muscle sections of one particular bait, DYSF, with its Y2H partners for which quality antibody was available for immuno-fluorescence assays. Among the 14 PPIs investigated, five showed no apparent colocalization while the others demonstrated different degrees of co-localization at the sarcolemma and/or in the cytoplasm (Figure 3). Analysis of Pearson's coefficient showed that DYSF labeling presented a strong correlation with the cardiomyopathy-associated protein 5 (CMYA5, 0.567), DES (0.656), filamin-C (FLNC, 0.521),kinesin family member 1B (KIF1B, 0.566) and optineurin (OPTN, 0.733), a medium correlation with Alstrom syndrome protein 1 (ALMS1, 0.402), diacylglycerol kinase delta (DGKD, 0.309), and nebulin (NEB, 0.332) and a low correlation with SNAPIN (0.12). We also visualized the sub-cellular localization of the interactions using a Duolink proximity ligation assay (PLA) for 44 PPIs. The Duolink technology is a combination of immunohistochemistry and rolling circle amplification and is based on the use of bifunctional probes consisting of a secondary antibody attached to a synthetic oligonucleotide. It generates a PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/16989806 quantifiable signal indicative of close proximity (<40 nm) between two antigens. For our purpose, primary antibodies against five different baits [DYSF, OPTN, -sarcoglycan (SGCG), adaptor protein containing PH domain, PTB domain and leucine zipper motif 1 (APPL1) and myomesin2 (MYOM2)] and some of their partners identified in our Y2H screenings (18 interacting partners for DYSF, 10 for OPTN, 3 for SGCG, 3 for APPL1 and 5 for MYOM2)ABI1-KTNACTN2-SNAPINDES-DYSFDYSF-APPLDYSF-KIF1BDYSF-SNAPINMYOM1-DYSFTCAP-ADPGKTCAP-KBTBDLeft lanes: InputMiddle lanes: ImmunoprecipitationRight lanes: negative controlFigure 2 Immunoprecipitation analysis of a subset of interactions. A subset of interactions from the baits: abl-interactor 1 (ABI1), DYSF, SNAP-associated protein (SNAPIN) and telethonin (TCAP) was assessed using co-immunoprecipitation using myogenic cells or.