Journal: Cells

Article Title: Co-Aggregation and Parallel Aggregation of Specific Proteins in Major Mental Illness

doi: 10.3390/cells12141848

Figure Lengend Snippet: Insoluble protein in human insular cortex samples, from 15 victims of suicide, 11 control individuals, 4 patients with MDD and 3 patients with Alzheimer’s disease. ( A – D ) Quantified levels of DISC1 ( A ), TRIOBP-1 ( B ), CRMP1 ( C ) and NPAS3 ( D ) protein seen in the insoluble protein fraction of post mortem insular cortex samples. Values are normalized to a common sample loaded on each membrane. Original Western blot data for DISC1, TRIOBP-1 and CRMP1 are in . NPAS3 data ( D ) have been published previously but are reanalyzed and summarized here for comparison. The NPAS3 data also include a few additional samples, which were not available for this study. ( E ) For the protein isoforms analyzed further (CRMP1 Lv and Sv, DISC1 70 kDa, TRIOBP-1 72 kDa, NPAS3 130 kDa), the proteins present in the insoluble pellet at a level at least 50% higher than the mean, which are interpreted as aggregating for the purposes of this study. The number of proteins found to be insoluble (aggregating) in each sample can be seen in . Graphs prepared using GraphPad Prism.

Article Snippet: Primary antibodies were purchased against β-actin (Origene, Rockville, MD, USA, OG-TA811000), CRMP1 (ProSci, Poway, CA, USA, 3625), DISC1 (Thermo Fisher Scientific, Weltham, MA, USA, 40-6800), Flag (Merck, Darmstadt, Germany, F1804), GFP (Merck, G6795) and TRIOBP-1 (Atlas Antibodies, Stockholm, Sweden, HPA019769).

Techniques: Control, Membrane, Western Blot, Comparison

Journal: Cells

Article Title: Co-Aggregation and Parallel Aggregation of Specific Proteins in Major Mental Illness

doi: 10.3390/cells12141848

Figure Lengend Snippet: Systematic pairwise testing of co-aggregation in cell culture. ( A ) Western blots of Flag-tagged proteins used in these experiments, expressed in HEK293 cells. ( B ) Equivalent blot of EGFP-fused proteins used in this experiment. Remaining images show constructs expressed in SH-SY5Y neuroblastoma cells: ( C ) Flag-tagged CRMP1 Sv, not aggregating. ( D ) Flag-tagged CRMP1 Lv, not aggregating. ( E ) Flag-tagged NPAS3, not aggregating. ( F ) Flag-tagged DISC1, aggregating. ( G ) Flag-tagged TRIOBP-1, aggregating. ( H ) Flag-tagged NPAS3 and EGFP-fused CRMP1 Sv, neither aggregating. ( I ) Flag-tagged NPAS3 and EGFP-fused CRMP1 Lv, neither aggregating. ( J ) Flag-tagged NPAS3 and EGFP-fused DISC1, only DISC1 is aggregating. ( K ) Flag-tagged NPAS3 and EGFP-fused TRIOBP-1, only TRIOBP-1 is aggregating. All cell photos are typical of 3 or more independent experiments. Scale bars represent 10 μm. shows versions of the experiments in ( H ) to ( K ) using the reciprocal vectors.

Article Snippet: Primary antibodies were purchased against β-actin (Origene, Rockville, MD, USA, OG-TA811000), CRMP1 (ProSci, Poway, CA, USA, 3625), DISC1 (Thermo Fisher Scientific, Weltham, MA, USA, 40-6800), Flag (Merck, Darmstadt, Germany, F1804), GFP (Merck, G6795) and TRIOBP-1 (Atlas Antibodies, Stockholm, Sweden, HPA019769).

Techniques: Cell Culture, Western Blot, Construct

Journal: Cells

Article Title: Co-Aggregation and Parallel Aggregation of Specific Proteins in Major Mental Illness

doi: 10.3390/cells12141848

Figure Lengend Snippet: Further systematic pairwise testing of co-aggregation in cell culture. Images show constructs co-expressed in SH-SY5Y neuroblastoma cells: ( A ) Flag-tagged TRIOBP-1 and EGFP-fused CRMP-1 Sv, only TRIOBP-1 aggregates. ( B ) Flag-tagged TRIOBP-1 and EGFP-fused CRMP-1 Lv, only TRIOBP-1 aggregates. ( C ) Flag-tagged DISC1 and EGFP-fused CRMP-1 Sv, example of cells, one with co-aggregation and one with aggregation of DISC1. ( D ) Flag-tagged DISC1 and EGFP-fused CRMP-1 Lv showing co-aggregation. ( E ) Flag-tagged CRMP1 Sv and EGFP-fused CRMP1 Lv showing co-aggregation. ( F ) Flag-tagged TRIOBP-1 and EGFP-fused DISC1, example of cells with co-aggregation and parallel aggregation. ( G ) Summary of co-aggregation results. All cell photos are typical of three or more independent experiments. Scale bars represent 10 μm. shows versions of these experiments using the reciprocal vectors.

Article Snippet: Primary antibodies were purchased against β-actin (Origene, Rockville, MD, USA, OG-TA811000), CRMP1 (ProSci, Poway, CA, USA, 3625), DISC1 (Thermo Fisher Scientific, Weltham, MA, USA, 40-6800), Flag (Merck, Darmstadt, Germany, F1804), GFP (Merck, G6795) and TRIOBP-1 (Atlas Antibodies, Stockholm, Sweden, HPA019769).

Techniques: Cell Culture, Construct

Journal: Cells

Article Title: Co-Aggregation and Parallel Aggregation of Specific Proteins in Major Mental Illness

doi: 10.3390/cells12141848

Figure Lengend Snippet: Investigating the co-aggregation of DISC1 and CRMP1. ( A , B ) Co-expression of Flag-DISC1 with EGFP, EGFP-CRMP1 Sv or EGFP-CRMP1 Lv in SH-SY5Y cells, analyzed by immunofluorescent microscopy in a blinded, quantified manner. For the purpose of this assay, an aggregate was defined as a punctate structure at least 1 µm in diameter. Results are an average of 10 coverslips per plasmid combination (9 in the case of DISC1 + EGFP), with 10 transfected cells examined per coverslip (or as many transfected cells as could be found). ( A ) Mean number of Flag (DISC1) aggregates per cell. ( B ) Mean size of DISC1 aggregates in cells displaying DISC1 aggregates. *: p < 0.05, ns: not significant, according to one-way ANOVA. ( C ) Western blots from an insoluble fraction purification assay. HEK293 cells were transfected with various combinations of Flag-tagged DISC1, CRMP1 Sv and Lv, lysed and then had their insoluble protein fraction purified. Samples are shown from both the unfractionated cell lysate and the purified insoluble fraction, and are representative of three independent experiments.

Article Snippet: Primary antibodies were purchased against β-actin (Origene, Rockville, MD, USA, OG-TA811000), CRMP1 (ProSci, Poway, CA, USA, 3625), DISC1 (Thermo Fisher Scientific, Weltham, MA, USA, 40-6800), Flag (Merck, Darmstadt, Germany, F1804), GFP (Merck, G6795) and TRIOBP-1 (Atlas Antibodies, Stockholm, Sweden, HPA019769).

Techniques: Expressing, Microscopy, Plasmid Preparation, Transfection, Western Blot, Purification

Journal: Nature communications

Article Title: Amino- and carboxyl-terminal domains of Filamin-A interact with CRMP1 to mediate Sema3A signalling.

doi: 10.1038/ncomms6325

Figure Lengend Snippet: Figure 3 | CRMP1 interacts with Filamin-A in vertebrates. (a) Interaction of Filamin-A and CRMPs in HEK293Tcells. HEK293Tcells were transfected with either one of CRMP1-5-V5 expression vector together with the Filamin-A-Myc vector or a mock vector. 72 h post transfection, cells were subjected to immunoprecipitation (IP) with an anti-Myc antibody. Immunoblotting was performed as indicated. Comparing with the faint bands of CRMP1-4 in the mock vector lanes, apparent V5 blots of CRMP1, CRMP3 and CRMP4 in the Filamin-A lanes indicate the physical interaction. (b) Interaction of Filamin- A and CRMP1 in rat E20 brain. The lysates from whole-brain of E20 rats were immunoprecipitated with anti-Filamin-A mouse mAb or control mouse IgG, then probed with an anti-CRMP1 antibody. Two CRMP1 bands represent the alternate splicing isoforms. Both forms of CRMP1 are co-immunoprecipitated with Filamin-A from rat brain. (c) Filamin-A mRNA expression in mouse E12 DRG and spinal cord. Left panel, antisense probe; right panel, sense probe. Filamin-A was expressed in DRG cell bodies (DRG) and motoneurons (MN). The neuroepithelial cell layer around the central canal in spinal cord (arrow in SpC) also expressed Filamin-A. Scale bar, 250 mm. (d) Co-localization of CRMP1, Filamin-A and F-actin in the Sema3A-stimulated DRG growth cones. Cultured mouse E12 DRG neurons were applied with or without Sema3A (0.5 U ml 1) for 5 min. The neurons were stained with anti-Filamin-A (blue) and anti-CRMP1 (red) antibodies and with Alexa488-conjugated Phalloidin (green). Without Sema3A stimulation, Filamin-A and CRMP1 were partially co-localized in the peripheral region of the growth cones. Sema3A stimulation induced the co-localization of CRMP1, Filamin-A and F-actin in the growth cones. Scale bar, 10 mm. (e,f) Suppression of Sema3A-induced collapse response of Filamin-A siRNA-transfected neurons. (e) The panels indicate the growth cone morphology with or without Sema3A (5 U ml 1) in mouse E12 DRG neurons transfected with Filamin-A or control siRNA. Growth cones were stained with Alexa488-Phalloidin. Scale bar, 10 mm. (f) Scored graph. White and black bars indicate no stimulation and Sema3A (5 U ml 1) stimulation, respectively. Each bar represents average±s.e.m. (n ¼ 5 from more than three independent cultures). **P ¼ 0.003, unpaired T-test.

Article Snippet: Mouse CRMP1 siRNA mixture (sc-60446) from Santa Cruz Biotech reduced CRMP1 expression in Neuro2A cells (Supplementary Fig. 2f).

Techniques: Transfection, Expressing, Plasmid Preparation, Immunoprecipitation, Western Blot, Control, Cell Culture, Staining

Journal: Nature communications

Article Title: Amino- and carboxyl-terminal domains of Filamin-A interact with CRMP1 to mediate Sema3A signalling.

doi: 10.1038/ncomms6325

Figure Lengend Snippet: Figure 4 | CRMP1 interacts with both amino- and carboxyl-terminal domains of Filamin-A. (a) Dot blot assay of Filamin-A fragments. Left panel, schematic representation of Filamin-A domains. Middle panel, dot blot analysis of Filamin-A fragments. A nitrocellurose membrane blotted with 5 mg of BSA or each Filamin-A fragment was incubated with recombinant CRMP1 (1 mg ml 1). Bound CRMP1 was visualized with an anti-CRMP1 antibody. CRMP1 bound to ABD and lesser extent to Ig6-10 and Ig21-24. Right panels, dot blot analysis of ABD and Ig21-24 subdomains. CH1, subdomain of ABD, and Ig24 bound to CRMP1. (b) Determination of CRMP1-binding residues in the ABD. One microgram of wild-type ABD and various ABD alanine mutants (S85A-K88A, M89A-K92A, H93A-R96A, T98A-Q101A, M102A-E105A) were spotted onto nitrocellulose membranes. The membranes were incubated with CRMP1-V5 and bound CRMP1 was detected by an anti-V5 antibody. ABD(H93A-R96A) mutant showed weak CRMP1 interaction. (c) Specific binding of Filamin-A wild type and Filamin-A(H93A-R96A) mutant to CRMP1. Various concentrations (0–0.6 mM) of Filamin-A were incubated with solid-phased CRMP1. Filamin-A(H93A-R96A) mutant (red line) binding is reduced compared with the Filamin-A wild type (black line) binding. Dissociation constants of wild type and H93A-R96A mutants are, respectively, estimated at 0.103 and 0.626 mM from this experiment with Scatchard analysis (inset). (d) Crystal structure of CH1 subdomain of ABD (3HOP). Yellow indicates Gln95 and Arg96. The side chains of these residues protrude from the molecular surface. Space-filled view of CH1 of ABD was rendered using Cn3D (version 4.3). (e) Morphology of chick E8 DRG growth cones expressing Filamin-A mutants. Control, wildtype, DABD, DIg1-15, DIg24 and Filamin-A(R96A) were stimulated with 3 U ml 1 Sema3A. Growth cones were stained with Alexa488- Phalloidin. (f) Quantification of Sema3A-induced collapse response. DABD and DIg24 significantly suppressed the response with Sema3A (3 U ml 1). ***P ¼ 0.0001, **P ¼ 0.003, unpaired T-test. (n ¼ 30, control; 33, wild type; 21, DABD; 10, DIg1-15; 13, DIg24 from more than three independent cultures). (g) Sema3A-growth cone collapse assay with Filamin-A(Q95A) and (R96A). Both single-alanine mutants significantly suppressed the growth cone collapse response with Sema3A (3 U ml 1) stimulation. *P ¼ 0.0272, unpaired T-test (n ¼ 9 from more than three independent cultures).

Article Snippet: Mouse CRMP1 siRNA mixture (sc-60446) from Santa Cruz Biotech reduced CRMP1 expression in Neuro2A cells (Supplementary Fig. 2f).

Techniques: Dot Blot, Membrane, Incubation, Recombinant, Binding Assay, Mutagenesis, Expressing, Control, Staining

Journal: Nature communications

Article Title: Amino- and carboxyl-terminal domains of Filamin-A interact with CRMP1 to mediate Sema3A signalling.

doi: 10.1038/ncomms6325

Figure Lengend Snippet: Figure 5 | Ile246 and Asn247 of CRMP1 are the key residues for Filamin-A interaction. (a) Binding of CRMP1 fragment 225–256 to the Filamin-A fragments. Other shorter fragments of CRMP1, 225–248 or 241–256, do not bind to the Filamin-A fragments. This indicates that the 225–256 fragment of CRMP1 comprises the minimal binding region to Filamin-A. Dashed circles indicate the spotted positions of the Filamin-A fragments. (b) Crystal structure of mouse CRMP1 (1KCX). Arrow indicates Arg245 to Asn247 residues (yellow) of CRMP1. Arrowhead indicates Gly490, a transition residue to the C-terminal region of CRMP1. Space-filled view of CRMP1 dimer was rendered using Cn3D. (c) ABD binding to CRMP1 mutants. Single-alanine mutants of CRMP1, R245A, I246A, N247A, but not C248A reduced the interaction with ABD. (d) Specific binding of Filamin-A wild type to CRMP1 or CRMP1(N247A). Various concentrations (0–1 mM) of Filamin-A were incubated with solid-phased CRMP1 wild type (black, solid line) or CRMP1(N247A) (red, dashed line). Dissociation constant of Filamin-A to CRMP1 wild type and CRMP1(N247A) were, respectively, estimated at 0.0787 and 0.191 mM from this experiment with Scatchard analysis (inset). (e) Micrograph of DRG growth cones expressing CRMP1 mutants with or without Sema3A stimulation (5 U ml 1). Growth cones were stained with Alexa488-Phalloidin. (f) Quantification of Sema3A-induced collapse response. I246A and N247A mutants suppressed Sema3A-induced growth cone collapse response. Data are mean±s.e.m. (n ¼ 10, control; 9, wildtype; 10, R245A; 10, I246A; N247A; 7, C248A from more than three independent cultures). *P ¼ 0.04, **P ¼ 0.0013, unpaired T-test.

Article Snippet: Mouse CRMP1 siRNA mixture (sc-60446) from Santa Cruz Biotech reduced CRMP1 expression in Neuro2A cells (Supplementary Fig. 2f).

Techniques: Binding Assay, Residue, Incubation, Expressing, Staining, Control

Journal: Nature communications

Article Title: Amino- and carboxyl-terminal domains of Filamin-A interact with CRMP1 to mediate Sema3A signalling.

doi: 10.1038/ncomms6325

Figure Lengend Snippet: Figure 6 | Ser522 phosphorylation regulates CRMP1–Fimain-A interaction. (a,b) Specific binding of CRMP1 wild type and CRMP1(S522D) to Filamin-A. (a) Binding curve. (b) Scatchard analysis. Various concentrations (0–3 mM) of CRMP1 were incubated with solid-phased Filamin-A. CRMP1(S522D) mutant (red line) binding is increased compared with CRMP1 wild type (black line) binding. Dissociation constants of wild type and S522D mutants were, respectively, estimated at 0.325 and 0.107 mM from this experiment. (c) Micrograph of chick E8 DRG growth cones expressing mock vector, CRMP1 wild type or CRMP1(S522D) with or without Sema3A stimulation (0.5 U ml 1). CRMP1(S522D) overexpression induced partially collapsed growth cone without Sema3A stimulation (left bottom). Growth cones were stained with Alexa488-Phalloidin. Scale bar, 10 mm. (d) Quantification of Sema3A-induced collapse response. CRMP1(S522D) significantly augmented Sema3A response. Data are mean±s.e.m. (none, n ¼ 10, control; 11, wild type; 6, S522D; Sema3A 0.5U, n ¼ 10, control; 10, wild type; 8, S522D; Sema3A 1U, n ¼ 8, control; 10, wild type; 6, S522D from more than three independent cultures). **Po0.01, unpaired T-test. (e,f) Suppression of CRMP1(S522D)-induced morphological alteration by Filamin-A knockdown. (e) Micrograph. Filamin-A RNAi prevented the mouse E13 DRG growth cones from the CRMP1(S522D)-induced morphological alteration. Growth cones were stained with Alexa488-Phalloidin. Scale bar, 10 mm. (f) Scored graph. Data are mean±s.e.m. (n ¼ 5 from more than three independent cultures). *Po0.05, unpaired T-test.

Article Snippet: Mouse CRMP1 siRNA mixture (sc-60446) from Santa Cruz Biotech reduced CRMP1 expression in Neuro2A cells (Supplementary Fig. 2f).

Techniques: Phospho-proteomics, Binding Assay, Incubation, Mutagenesis, Expressing, Plasmid Preparation, Over Expression, Staining, Control, Knockdown

Journal: Nature communications

Article Title: Amino- and carboxyl-terminal domains of Filamin-A interact with CRMP1 to mediate Sema3A signalling.

doi: 10.1038/ncomms6325

Figure Lengend Snippet: Figure 7 | CRMP1(S522D) induces morphological alternation of Filamin-A. (a) Co-sedimentation assay of CRMP1(S522D) and Filamin-A. Filamin-A (0.03–1 mM) was reconstituted with or without 1 mM CRMP1(S522D) and incubated at 25 C for 1 h then ultracentrifuged. Separated specimens were subjected to 7.5% polyacrylamide gel electrophoresis and stained with Coomassie Brilliant Blue. An arrow (280 kDa) and three arrowheads (64, 60, 55 kDa) indicate Filamin-A and CRMP1(S522D), respectively. The highest CRMP1 band (64 kDa) and two lower fragments (60 and 55 kDa) represent full-length and partially cleaved products, respectively. The 66-kDa band (*) is contaminant from E. coli. S ¼ supernatant, P ¼ precipitate, M ¼ protein size marker. (b) Filamin-A binding curve and Scatchard plot. The specific binding is represented as the molar ratio of Filamin-A/CRMP1(S522D). Each point represents the average±s.e.m. of three to four independent values. Scatchard plot (inset) indicates that Filamin-A possesses high- and low-affinity binding sites of which dissociation constants are 0.131 and 0.78 mM, respectively. Stoichiometry of the high- and low-affinity binding sites is estimated as 0.065 and 0.227 (Filamin-A/CRMP1(S522D) molar ratio), respectively. (c–k) Atomic morphology and height profile of purified proteins on the sapphire (0001) surface. (c–e) Filamin-A (10 nM); (f–h) CRMP1(S522D) (40 nM); (i–k) Filamin-A (10 nM) and CRMP1(S522D) (20 nM) mixture. The specimens were scanned with Seiko Instruments Atomic force microscopy equipped with a cantilever (SI-DF3, radius 10 nm, spring constant 1.6 N ml 1) in tapping mode. (c) The Filamin-A specimen was scanned 3 3 mm2 area and its partial view is shown. The ladders on the sapphire surfaces are atomic steps. White arrows indicate the typical molecular shape of Filamin-A. (f,i) CRMP1(S522D) and Filamin-A-CRMP1(S522D) mixture were scanned 1 1 mm2

Article Snippet: Mouse CRMP1 siRNA mixture (sc-60446) from Santa Cruz Biotech reduced CRMP1 expression in Neuro2A cells (Supplementary Fig. 2f).

Techniques: Sedimentation, Incubation, Polyacrylamide Gel Electrophoresis, Staining, Marker, Binding Assay, Microscopy

Journal: Nature communications

Article Title: Amino- and carboxyl-terminal domains of Filamin-A interact with CRMP1 to mediate Sema3A signalling.

doi: 10.1038/ncomms6325

Figure Lengend Snippet: Figure 8 | Sema3A stimulation reduces the co-sedimentation of Filamin-A with F-actin from HEK293 cells. (a) Falling ball viscometry. The gelation of F-actin and Filamin-A with CRMP1 was examined. While addition of CRMP1 wild type did not affect the strength of Filamin-F-actin gel (solid line), CRMP1(S522D) reduced the gel viscosity and the beads placed on the gel fell by gravity (dashed line). Data are mean±s.e.m. (n ¼ 6). **P ¼ 0.0044, unpaired T-test. (b,c) Co-sedimentation of Filamin-A and F-actin from HEK293 cells expressing Filamin-A and CRMP1 mutants. Cell lysates were ultracentrifuged to precipitate F-actin and its associated proteins. Expression of CRMP1 wild type (wt) but not CRMP1(N247A) enhanced the co- precipitation of Filamin-A. (c) Scored graph. Data are mean±s.e.m. (n ¼ 6 from more than five independent experiments). *P ¼ 0.036, unpaired T-test. (d) Time course of Filamin-A co-sedimentation after Sema3A stimulation. HEK293 cells expressing Filamin-A-myc, CRMP1 wild type, HA-NRP1 and HA-PlexA1 were stimulated with Sema3A for (8 U ml 1) 0 to 10 min. Co-sedimentation of Filamin-A was decreased after 10 min Sema3A stimulation. (e,f) Attenuation by mutant proteins. HEK293 cells expressing Filamin-A, CRMP1, NRP1 and PlexA1 were stimulated with Sema3A (8 U ml 1) for 10 min. In the CRMP1 wild type (wt)-transfected cells, Sema3A stimulation reduced co-sedimentation of Filamin-A B50% compared with the un-stimulated cells. In contrast, the cells expressing CRMP1(N247A) or Filamin-A(R96A) did not alter the Filamin-A precipitation on Sema3A stimulation. (f) Scored graph. Data are mean±s.e.m. (n ¼ 14, wild type; 10, CRMP1(N247A); 9, Filamin-A(R96A) from more than five independent experiments). **P ¼ 0.0025, unpaired T-test.

Article Snippet: Mouse CRMP1 siRNA mixture (sc-60446) from Santa Cruz Biotech reduced CRMP1 expression in Neuro2A cells (Supplementary Fig. 2f).

Techniques: Sedimentation, Viscometric Assay, Viscosity, Expressing, Mutagenesis, Transfection

Journal: Nature communications

Article Title: Amino- and carboxyl-terminal domains of Filamin-A interact with CRMP1 to mediate Sema3A signalling.

doi: 10.1038/ncomms6325

Figure Lengend Snippet: Figure 9 | Schematic representation of CRMP1–Filamin-A interaction in Sema3A signalling. (a,b) Two possible configurations of CRMP1 and Filamin-A complex. (a) One CRMP1 tetramer binds to either the pair of N-terminal actin-binding domain (ABD) or the pair of the C-terminal Ig24. (b) One CRMP1 tetramer simultaneously binds to ABD and Ig24. (c) Sema3A–CRMP1 signalling. Sema3A stimulation activates Cdk5, which subsequently phosphorylates the Ser522 of CRMP1. This phosphorylation facilitates the interaction of CRMP1 and Filamin-A. (d) Crosslinking F-actin by Filamin-A. Filamin-A dimer binds to F-actin through the actin-binding domain (ABD) of each branch. This brings F-actin crosslinking. (e) Removal of Filamin-A from F-actin. Formation of Filamin-A and CRMP1 complex would remove Filamin-A from F-actin in turn to weaken the cytoskeleton.

Article Snippet: Mouse CRMP1 siRNA mixture (sc-60446) from Santa Cruz Biotech reduced CRMP1 expression in Neuro2A cells (Supplementary Fig. 2f).

Techniques: Binding Assay, Phospho-proteomics

Journal: PLoS ONE

Article Title: Differentially expressed proteins underlying childhood cortical dysplasia with epilepsy identified by iTRAQ proteomic profiling

doi: 10.1371/journal.pone.0172214

Figure Lengend Snippet: 64 up-regulated proteins in childhood cortical dysplasia patients with epilepsy compared to controls by iTRAQ.

Article Snippet: The sequences of primers (OriGene Technologies, Inc. Rockville, USA) were NDRG1 (HP209104), FSCN1 (HP206673), FABP3 (HP207465), DPYSL5 (HP213501), CRMP1 (HP232913), PRDX6 (HP208150), PSAP (HP231407), MAP4 (HP206072), and β-Actin (HP204660).

Techniques: Sequencing, Variant Assay

Journal: PLoS ONE

Article Title: Differentially expressed proteins underlying childhood cortical dysplasia with epilepsy identified by iTRAQ proteomic profiling

doi: 10.1371/journal.pone.0172214

Figure Lengend Snippet: Quantification of protein levels showed increased expression of CRMP1 (A), DPYSL5 (B), NDRG1 (C) and FSCN1 (D) and decreased expression of PRDX6 (E) in brain tissue of childhood cortical dysplasia patients compared to controls. *p<0.05 compared to controls.

Article Snippet: The sequences of primers (OriGene Technologies, Inc. Rockville, USA) were NDRG1 (HP209104), FSCN1 (HP206673), FABP3 (HP207465), DPYSL5 (HP213501), CRMP1 (HP232913), PRDX6 (HP208150), PSAP (HP231407), MAP4 (HP206072), and β-Actin (HP204660).

Techniques: Expressing

Journal: PLoS ONE

Article Title: Differentially expressed proteins underlying childhood cortical dysplasia with epilepsy identified by iTRAQ proteomic profiling

doi: 10.1371/journal.pone.0172214

Figure Lengend Snippet: IHC score of CRMP1 (A & B), DPYSL5 (C & D) and FSCN1 (E & F) were significantly increased in brain tissues of childhood cortical dysplasia patients with epilepsy compared to controls. *p<0.05 compared to controls.

Article Snippet: The sequences of primers (OriGene Technologies, Inc. Rockville, USA) were NDRG1 (HP209104), FSCN1 (HP206673), FABP3 (HP207465), DPYSL5 (HP213501), CRMP1 (HP232913), PRDX6 (HP208150), PSAP (HP231407), MAP4 (HP206072), and β-Actin (HP204660).

Techniques: