Journal: Archives of Medical Science : AMS

Article Title: Tissue expression of β-catenin and E- and N-cadherins in chronic hepatitis C and hepatocellular carcinoma

doi: 10.5114/aoms.2017.65272

Figure Lengend Snippet: Detection of β-catenin, E- and N-cadherins and the cellular localization of the proteins in chronic hepatitis C (CH-C), hepatocellular carcinoma (HCC) and normal liver (control)

Article Snippet: Mouse anti-human monoclonal antibodies (mAbs) were employed, directed against human beta-catenin (Clone 196618; Code M 7052) (in dilution 1 : 50) (R&D Systems, UK), against E-cadherin (clone NCH-38) (in dilution 1 : 100) (DakoCytomation, Gdynia, PL) and against N-cadherin (in dilution 1 : 50) (clone IAR06) (Novocastra Labs. Ltd, Newcastle upon Tyne, UK).

Techniques: Control

Journal: Archives of Medical Science : AMS

Article Title: Tissue expression of β-catenin and E- and N-cadherins in chronic hepatitis C and hepatocellular carcinoma

doi: 10.5114/aoms.2017.65272

Figure Lengend Snippet: Immunohistochemical ( A – C ) and hybridocytochemical localization of β-catenin ( D ) in liver. Membranous/ cytoplasmic localization of β-catenin in liver with chronic hepatitis C ( A ), membranous localization of β-catenin in hepatocellular carcinoma ( B ) and in control liver ( C ); mRNA for β-catenin in cytoplasm and cell nuclei in HCC fragment ( D ). Immunocytochemical detection of E-cadherin ( E – F ) and N-cadherin ( H ) in liver. Predominantly membranous localization of E-cadherin in liver with chronic hepatitis C ( E ), hepatocellular carcinoma ( F ) and control liver ( G ). Membrano-cytoplasmic localization of N-cadherin in fragment of hepatocellular carcinoma ( H ). Immunohistochemistry ( A – C , E – H ) technique and hybridization in situ method ( D ). Hematoxylin counterstained. Bar = 40 μm

Article Snippet: Mouse anti-human monoclonal antibodies (mAbs) were employed, directed against human beta-catenin (Clone 196618; Code M 7052) (in dilution 1 : 50) (R&D Systems, UK), against E-cadherin (clone NCH-38) (in dilution 1 : 100) (DakoCytomation, Gdynia, PL) and against N-cadherin (in dilution 1 : 50) (clone IAR06) (Novocastra Labs. Ltd, Newcastle upon Tyne, UK).

Techniques: Immunohistochemical staining, Control, Immunohistochemistry, Hybridization, In Situ

Journal: Archives of Medical Science : AMS

Article Title: Tissue expression of β-catenin and E- and N-cadherins in chronic hepatitis C and hepatocellular carcinoma

doi: 10.5114/aoms.2017.65272

Figure Lengend Snippet: Comparison of quantitatively assessed β-catenin, E-cadherin and N-cadherin immunoexpression (% of the IHC reaction area in analysed area of liver parenchyma) in chronic hepatitis C (CH-C), hepatocellular carcinoma (HCC) and normal liver (C)

Article Snippet: Mouse anti-human monoclonal antibodies (mAbs) were employed, directed against human beta-catenin (Clone 196618; Code M 7052) (in dilution 1 : 50) (R&D Systems, UK), against E-cadherin (clone NCH-38) (in dilution 1 : 100) (DakoCytomation, Gdynia, PL) and against N-cadherin (in dilution 1 : 50) (clone IAR06) (Novocastra Labs. Ltd, Newcastle upon Tyne, UK).

Techniques: Comparison

Journal: Archives of Medical Science : AMS

Article Title: Tissue expression of β-catenin and E- and N-cadherins in chronic hepatitis C and hepatocellular carcinoma

doi: 10.5114/aoms.2017.65272

Figure Lengend Snippet: Comparative immunoexpression of β-catenin, E-cadherin and N-cadherin in liver with chronic hepatitis C (CH-C), hepatocellular carcinoma (HCC) and normal organ (control) ***p (level of significance) value < 0.001, *p < 0.05.

Article Snippet: Mouse anti-human monoclonal antibodies (mAbs) were employed, directed against human beta-catenin (Clone 196618; Code M 7052) (in dilution 1 : 50) (R&D Systems, UK), against E-cadherin (clone NCH-38) (in dilution 1 : 100) (DakoCytomation, Gdynia, PL) and against N-cadherin (in dilution 1 : 50) (clone IAR06) (Novocastra Labs. Ltd, Newcastle upon Tyne, UK).

Techniques: Control

Journal: Archives of Medical Science : AMS

Article Title: Tissue expression of β-catenin and E- and N-cadherins in chronic hepatitis C and hepatocellular carcinoma

doi: 10.5114/aoms.2017.65272

Figure Lengend Snippet: Tissue expression of β-catenin, E-cadherin and N-cadherin (mean % of IHC reaction area ± SD) as related to grading and staging in chronic hepatitis C (CH-C) group

Article Snippet: Mouse anti-human monoclonal antibodies (mAbs) were employed, directed against human beta-catenin (Clone 196618; Code M 7052) (in dilution 1 : 50) (R&D Systems, UK), against E-cadherin (clone NCH-38) (in dilution 1 : 100) (DakoCytomation, Gdynia, PL) and against N-cadherin (in dilution 1 : 50) (clone IAR06) (Novocastra Labs. Ltd, Newcastle upon Tyne, UK).

Techniques: Expressing

Journal: Archives of Medical Science : AMS

Article Title: Tissue expression of β-catenin and E- and N-cadherins in chronic hepatitis C and hepatocellular carcinoma

doi: 10.5114/aoms.2017.65272

Figure Lengend Snippet: Values of Spearman’s rank coefficient between expression of β-catenin, E-cadherin, N-cadherin (% IHC reaction area per area of hepatic parenchyma) and clinicopathological data in CH-C group

Article Snippet: Mouse anti-human monoclonal antibodies (mAbs) were employed, directed against human beta-catenin (Clone 196618; Code M 7052) (in dilution 1 : 50) (R&D Systems, UK), against E-cadherin (clone NCH-38) (in dilution 1 : 100) (DakoCytomation, Gdynia, PL) and against N-cadherin (in dilution 1 : 50) (clone IAR06) (Novocastra Labs. Ltd, Newcastle upon Tyne, UK).

Techniques: Expressing, Infection

Journal: Heliyon

Article Title: Cortical neurons obtained from patient-derived iPSCs with GNAO1 p.G203R variant show altered differentiation and functional properties

doi: 10.1016/j.heliyon.2024.e26656

Figure Lengend Snippet: Analysis of neuronal rosettes morphology in the independent GNAO1 +/G203R #2 line A. Phase contrast images of GNAO1 +/G203R #2 cells at the neural rosette stage, corresponding to day 30 of differentiation. Note that the in the protocol used for differentiation of GNAO1 +/G203R #2 cells (as described in the Methods section) has a different timing from the one depicted in A. B-D. Immunostaining was performed using the specified primary antibodies along with DAPI for nuclear labeling. TBR1 is a marker of early-born neurons (B); and TUJ1 (B) and MAP2 (C) are markers for neurons; NESTIN serves as a marker for neural progenitors (C); β-catenin as a marker of apical polarization in neural rosettes (D). Number of individual cultures for experiments was 3.

Article Snippet: Cells were then incubated overnight at 4 °C with primary antibodies at the following dilutions: rabbit anti-MAP2 (ab254264 Abcam, 1:2000), mouse anti-βTUBULIN III (TUJ1) (T8660 Sigma-Aldrich, 1:500), mouse Anti-Nestin (ab6142 Abcam, 1:250), rabbit anti-TBR1 (ab183032 Abcam,1:200), mouse anti-β-Catenin (571–781 BD Transduction Laboratories, 1:200).

Techniques: Immunostaining, Labeling, Marker

Journal: The Journal of Neuroscience

Article Title: Calcineurin Inhibition Causes α2δ-1–Mediated Tonic Activation of Synaptic NMDA Receptors and Pain Hypersensitivity

doi: 10.1523/JNEUROSCI.0282-20.2020

Figure Lengend Snippet: Treatment with FK506 increases α2δ-1–NMDAR interaction and synaptic trafficking in the spinal cord. A, B, Original blotting images (A, two pairs of samples) and quantification (B) of α2δ-1 protein levels in the DRG and dorsal spinal cord tissues of vehicle-treated (veh) and FK506-treated (FK) mice (n = 8 mice per group). C, D, Representative blotting image (C) and mean data (D) show the interaction between α2δ-1 and GluN1 in the tissue extracts of dorsal spinal cords from mice treated with vehicle and FK506 (n = 8 mice per group). Proteins were immunoprecipitated (IP) initially with a rabbit anti-GluN1 or anti-IgG antibody. Immunoblotting was then performed using mouse anti-α2δ-1 or rabbit anti-GluN1 antibodies. IgG and input (tissue lysates only, without immunoprecipitation) were used as negative and positive controls, respectively. E, F, Representative gel images (E, two pairs of samples) and quantification (F) of the protein levels of GluN1, α2δ-1, and PSD-95 (a synaptic marker) in synaptosomes isolated from dorsal spinal cord tissues of vehicle-treated and FK506-treated mice (n = 9 mice per group). GAPDH or PSD-95 was used as the internal control for normalizing the protein level on the same gel. M, Molecular weight marker. B, D, F, Values are normalized to expression levels in vehicle-treated mice in each group. Data are mean ± SEM. *p < 0.05; **p < 0.01; ***p < 0.001; compared with vehicle-treated group (two-tailed Student's t test).

Article Snippet: The membrane was treated with 5% blotting-grade buffer in TBS with Tween 20 for 1 h at 25°C and then incubated with the following primary antibodies overnight at 4°C: mouse anti-α2δ-1 (#sc-271697, 1:1000; Santa Cruz Biotechnology), rabbit anti-GluN1 (#G-8913, 1:1000; Sigma Millipore), or rabbit anti-GAPDH (#5174, 1:2000; Cell Signaling Technology).

Techniques: Immunoprecipitation, Western Blot, Marker, Isolation, Molecular Weight, Expressing, Two Tailed Test

Journal: The Journal of Neuroscience

Article Title: Calcineurin Inhibition Causes α2δ-1–Mediated Tonic Activation of Synaptic NMDA Receptors and Pain Hypersensitivity

doi: 10.1523/JNEUROSCI.0282-20.2020

Figure Lengend Snippet: α2δ-1 is essential for FK506-induced activation of presynaptic NMDARs in the spinal dorsal horn. A, B, Original recording traces (A) and cumulative probability plots (B) show the effect of bath application of 50 μm AP5 on mEPSCs of lamina II neurons from vehicle-treated mice, FK506-treated mice, FK506-treated mice from which spinal cord slices were incubated with 100 μm gabapentin, and FK506-treated Cacna2d1-KO mice. C, D, Summary data show the effect of AP5 on the frequency (C) and amplitude (D) of mEPSCs of lamina II neurons from vehicle-treated mice (n = 12 neurons from 4 mice), FK506-treated mice (n = 11 neurons from 5 mice), FK506-treated mice from which spinal cord slices were incubated with gabapentin (n = 13 neurons from 6 mice), and FK506-treated Cacna2d1-KO mice (n = 12 neurons from 4 mice). Data are mean ± SEM. *p < 0.05 compared with respective baseline (one-way ANOVA followed by Tukey's post hoc test). ##p < 0.01 compared with baseline in the vehicle group (one-way ANOVA followed by Tukey's post hoc test).

Article Snippet: The membrane was treated with 5% blotting-grade buffer in TBS with Tween 20 for 1 h at 25°C and then incubated with the following primary antibodies overnight at 4°C: mouse anti-α2δ-1 (#sc-271697, 1:1000; Santa Cruz Biotechnology), rabbit anti-GluN1 (#G-8913, 1:1000; Sigma Millipore), or rabbit anti-GAPDH (#5174, 1:2000; Cell Signaling Technology).

Techniques: Activation Assay, Incubation

Journal: The Journal of Neuroscience

Article Title: Calcineurin Inhibition Causes α2δ-1–Mediated Tonic Activation of Synaptic NMDA Receptors and Pain Hypersensitivity

doi: 10.1523/JNEUROSCI.0282-20.2020

Figure Lengend Snippet: α2δ-1 mediates FK506-induced activation of NMDARs at primary afferent terminals in the spinal dorsal horn. A, B, Original recording traces (A) and summary data (B) show the effect of 50 μm AP5 on the amplitude of evoked monosynaptic EPSCs of lamina II neurons from vehicle-treated mice (n = 10 neurons from 4 mice), FK506-treated mice (n = 9 neurons from 5 mice), FK506-treated mice from which spinal cord slices were incubated with 100 μm gabapentin (n = 12 neurons from 6 mice), and FK506-treated Cacna2d1-KO mice (n = 12 neurons from 4 mice). C, D, Representative recording traces (C) and mean data (D) show the effect of 50 μm AP5 on the PPR of evoked EPSCs on lamina II neurons from vehicle-treated mice (n = 10 neurons from 4 mice), FK506-treated mice (n = 8 neurons from 5 mice), FK506-treated mice from which spinal cord slices were incubated with gabapentin (n = 13 neurons from 6 mice), and FK506-treated Cacna2d1-KO mice (n = 11 neurons from 4 mice). Data are mean ± SEM. *p < 0.05; **p < 0.01; compared with respective baseline; #p < 0.05 compared with baseline in the vehicle group (one-way ANOVA followed by Tukey's post hoc test).

Article Snippet: The membrane was treated with 5% blotting-grade buffer in TBS with Tween 20 for 1 h at 25°C and then incubated with the following primary antibodies overnight at 4°C: mouse anti-α2δ-1 (#sc-271697, 1:1000; Santa Cruz Biotechnology), rabbit anti-GluN1 (#G-8913, 1:1000; Sigma Millipore), or rabbit anti-GAPDH (#5174, 1:2000; Cell Signaling Technology).

Techniques: Activation Assay, Incubation

Journal: The Journal of Neuroscience

Article Title: Calcineurin Inhibition Causes α2δ-1–Mediated Tonic Activation of Synaptic NMDA Receptors and Pain Hypersensitivity

doi: 10.1523/JNEUROSCI.0282-20.2020

Figure Lengend Snippet: α2δ-1-bound NMDARs are involved in FK506-induced activation of presynaptic NMDARs in the spinal dorsal horn. A, B, Original recording traces (A) and cumulative probability plots (B) show the effect of bath application of 50 μm AP5 on mEPSCs of lamina II neurons from FK506-treated WT mice in which spinal cord slices were incubated with 1 μm control peptide or 1 μm α2δ-1Tat peptide. C, D, Summary data show the effect of AP5 on the frequency (C) and amplitude (D) of mEPSCs of lamina II neurons from FK506-treated WT mice in which spinal cord slices were incubated with 1 μm control peptide (n = 10 neurons from 6 mice) or 1 μm α2δ-1Tat peptide (n = 10 neurons from 5 mice). Data are mean ± SEM. *p < 0.05 compared with respective baseline; ###p < 0.001 compared with baseline in the control peptide group (one-way ANOVA followed by Tukey's post hoc test).

Article Snippet: The membrane was treated with 5% blotting-grade buffer in TBS with Tween 20 for 1 h at 25°C and then incubated with the following primary antibodies overnight at 4°C: mouse anti-α2δ-1 (#sc-271697, 1:1000; Santa Cruz Biotechnology), rabbit anti-GluN1 (#G-8913, 1:1000; Sigma Millipore), or rabbit anti-GAPDH (#5174, 1:2000; Cell Signaling Technology).

Techniques: Activation Assay, Incubation

Journal: The Journal of Neuroscience

Article Title: Calcineurin Inhibition Causes α2δ-1–Mediated Tonic Activation of Synaptic NMDA Receptors and Pain Hypersensitivity

doi: 10.1523/JNEUROSCI.0282-20.2020

Figure Lengend Snippet: α2δ-1-bound NMDARs mediate FK506 treatment-induced activation of NMDARs at primary afferent terminals in the spinal dorsal horn. A, B, Representative current traces (A) and mean data (B) show the effect of 50 μm AP5 on the amplitude of monosynaptically evoked EPSCs of lamina II neurons from FK506-treated WT mice from which spinal cord slices were incubated with 1 μm control peptide (n = 12 neurons from 6 mice) or 1 μm α2δ-1Tat peptide (n = 10 neurons from 5 mice). C, D, Original recording traces (C) and summary data (D) show the effect of 50 μm AP5 on the PPR of evoked EPSCs on lamina II neurons from FK506-treated WT mice from which spinal cord slices were incubated with 1 μm control peptide (n = 9 neurons from 6 mice) or 1 μm α2δ-1Tat peptide (n = 9 neurons from 5 mice). Data are mean ± SEM. *p < 0.05; **p < 0.01; compared with respective baseline. ##p < 0.01; ###p < 0.001; compared with baseline in the control peptide group (one-way ANOVA followed by Tukey's post hoc test).

Article Snippet: The membrane was treated with 5% blotting-grade buffer in TBS with Tween 20 for 1 h at 25°C and then incubated with the following primary antibodies overnight at 4°C: mouse anti-α2δ-1 (#sc-271697, 1:1000; Santa Cruz Biotechnology), rabbit anti-GluN1 (#G-8913, 1:1000; Sigma Millipore), or rabbit anti-GAPDH (#5174, 1:2000; Cell Signaling Technology).

Techniques: Activation Assay, Incubation

Journal: The Journal of Neuroscience

Article Title: Calcineurin Inhibition Causes α2δ-1–Mediated Tonic Activation of Synaptic NMDA Receptors and Pain Hypersensitivity

doi: 10.1523/JNEUROSCI.0282-20.2020

Figure Lengend Snippet: α2δ-1 is required for FK506-induced increases in postsynaptic NMDAR activity in the spinal dorsal horn. A, B, Representative current traces (A) and mean data (B) show NMDAR currents elicited by puff application of 100 μm NMDA onto lamina II neurons from vehicle-treated WT mice (n = 11 neurons from 4 mice), FK506-treated WT mice (n = 11 neurons from 5 mice), FK506-treated Cacna2d1-KO mice (n = 10 neurons from 4 mice), and FK506-treated mice from which spinal cord slices were incubated with 100 μm gabapentin (n = 9 neurons from 4 mice). C, D, Original recording traces (C) and group data (D) show puff NMDA-elicited currents in lamina II neurons from FK506-treated mice from which spinal cord slices were incubated with 1 μm control peptide (n = 12 neurons from 5 mice) or 1 μm α2δ-1Tat peptide (n = 10 neurons from 4 mice). Data are mean ± SEM. *p < 0.05; **p < 0.01; compared with the vehicle or control peptide group (one-way ANOVA followed by Tukey's post hoc test in B; two-tailed Student's t test in D).

Article Snippet: The membrane was treated with 5% blotting-grade buffer in TBS with Tween 20 for 1 h at 25°C and then incubated with the following primary antibodies overnight at 4°C: mouse anti-α2δ-1 (#sc-271697, 1:1000; Santa Cruz Biotechnology), rabbit anti-GluN1 (#G-8913, 1:1000; Sigma Millipore), or rabbit anti-GAPDH (#5174, 1:2000; Cell Signaling Technology).

Techniques: Activity Assay, Incubation, Two Tailed Test

Journal: The Journal of Neuroscience

Article Title: Calcineurin Inhibition Causes α2δ-1–Mediated Tonic Activation of Synaptic NMDA Receptors and Pain Hypersensitivity

doi: 10.1523/JNEUROSCI.0282-20.2020

Figure Lengend Snippet: α2δ-1 and NMDARs expressed in primary sensory neurons contribute to FK506-induced pain hypersensitivity. A, Time course of changes in paw withdrawal thresholds tested with von Frey filaments, a heat stimulus, and a noxious pressure stimulus in WT and Cacna2d1-KO mice treated with FK506 (n = 11 mice per group) and in Grin1-cKO mice treated with vehicle or FK506 (n = 10 mice per group) for 7 d. Solid bar represents daily drug injections. Data are mean ± SEM. *p < 0.05; **p < 0.01; ***p < 0.001; compared with respective baselines before treatment with FK506 (time 0). #p < 0.05; ##p < 0.01; ###p < 0.001; compared with the WT+FK506 group at the same time point (two-way ANOVA followed by Dunnett's or Tukey's post hoc test). B, C, Representative blotting images (B) and mean data (C) show the GluN1 protein levels in the DRG and dorsal spinal cord tissues from WT and Grin1-cKO mice (n = 11 mice per group). M, Molecular weight marker. Data are mean ± SEM. ***p < 0.001 compared with the WT group (two-tailed Student's t test).

Article Snippet: The membrane was treated with 5% blotting-grade buffer in TBS with Tween 20 for 1 h at 25°C and then incubated with the following primary antibodies overnight at 4°C: mouse anti-α2δ-1 (#sc-271697, 1:1000; Santa Cruz Biotechnology), rabbit anti-GluN1 (#G-8913, 1:1000; Sigma Millipore), or rabbit anti-GAPDH (#5174, 1:2000; Cell Signaling Technology).

Techniques: Molecular Weight, Marker, Two Tailed Test

Journal: The Journal of Neuroscience

Article Title: Calcineurin Inhibition Causes α2δ-1–Mediated Tonic Activation of Synaptic NMDA Receptors and Pain Hypersensitivity

doi: 10.1523/JNEUROSCI.0282-20.2020

Figure Lengend Snippet: α2δ-1 inhibition or KO and blocking α2δ-1-NMDAR interaction reverse FK506-induced pain hypersensitivity in mice. A, Time course of the effect of intraperitoneal injection with 60 mg/kg gabapentin (GBP) on paw withdrawal thresholds tested with von Frey filaments, a heat stimulus, and a noxious pressure stimulus in FK506-treated WT mice and Cacna2d1-KO mice (n = 8 mice per group). B, Time course of the effects of intrathecal injection with 1 μg control peptide or 1 μg α2δ-1Tat peptide on paw withdrawal thresholds in response to von Frey filaments, a heat stimulus, and a noxious pressure stimulus in FK506-treated WT mice and FK506-treated Cacna2d1-KO mice (n = 10 mice per group). Data are mean ± SEM. *p < 0.05; **p < 0.01; ***p < 0.001; compared with the baseline before drug injection (time 0; repeated-measures ANOVA followed by Dunnett's post hoc test).

Article Snippet: The membrane was treated with 5% blotting-grade buffer in TBS with Tween 20 for 1 h at 25°C and then incubated with the following primary antibodies overnight at 4°C: mouse anti-α2δ-1 (#sc-271697, 1:1000; Santa Cruz Biotechnology), rabbit anti-GluN1 (#G-8913, 1:1000; Sigma Millipore), or rabbit anti-GAPDH (#5174, 1:2000; Cell Signaling Technology).

Techniques: Inhibition, Blocking Assay, Injection