Journal: International Journal of Molecular Medicine

Article Title: Activating metabotropic glutamate receptor-7 attenuates visceral hypersensitivity in neonatal maternally separated rats

doi: 10.3892/ijmm.2018.4022

Figure Lengend Snippet: Sequences of primers used for reverse transcription-quantitative polymerase chain reaction.

Article Snippet: Tissues were incubated overnight at 4°C with anti-mGluR7 antibody (1:500, rabbit anti-rat, cat. no. NB-91787; Novus Biologicals LLC), or antibodies against cluster of differentiation (CD)3 (1:100, rabbit anti-rat, cat. no. ab16669) CD68 (1:200, mouse anti-rat, cat. no. ab31630) or myeloperoxidase (MPO; 1:50, rabbit anti-rat, cat. no. ab9535) (Abcam, Cambridge, UK).

Techniques:

Journal: International Journal of Molecular Medicine

Article Title: Activating metabotropic glutamate receptor-7 attenuates visceral hypersensitivity in neonatal maternally separated rats

doi: 10.3892/ijmm.2018.4022

Figure Lengend Snippet: mGluR7 expression in the colon of rats in the NC and NMS groups. (A) Immunohistochemistry of mGluR7 in rat colons. Staining in the mucosa (black arrowheads) was compared between the NC and NMS group; there was no difference beyond the submucosa (magnification, ×200). (B) mRNA expression levels of mGluR7 in the rat colons. (C) Protein expression levels of mGluR7 in the rat colons. (D) Grey values were used in the semi-quantitative analysis of western blotting results; mGluR7/β-actin ratio was used to indicate relative protein expression levels of mGluR7. Data were analysed by Student's t-test and are expressed as the means ± standard error of the mean (number of rats=6). * P<0.05. mGluR7, metabotropic glutamate receptor-7; NC, normal control; NMS, neonatal maternal separation.

Article Snippet: Tissues were incubated overnight at 4°C with anti-mGluR7 antibody (1:500, rabbit anti-rat, cat. no. NB-91787; Novus Biologicals LLC), or antibodies against cluster of differentiation (CD)3 (1:100, rabbit anti-rat, cat. no. ab16669) CD68 (1:200, mouse anti-rat, cat. no. ab31630) or myeloperoxidase (MPO; 1:50, rabbit anti-rat, cat. no. ab9535) (Abcam, Cambridge, UK).

Techniques: Expressing, Immunohistochemistry, Staining, Western Blot, Control

Journal: International Journal of Molecular Medicine

Article Title: Activating metabotropic glutamate receptor-7 attenuates visceral hypersensitivity in neonatal maternally separated rats

doi: 10.3892/ijmm.2018.4022

Figure Lengend Snippet: Effects of AMN082 on visceral hypersensitivity in NMS rats. AMN082 (3 or 10 mg/kg) was administered intraperitoneally 1 h prior to the tests. (A) Protein expression levels of mGluR7 in the colon samples obtained from rats in the NMS and NMS + AMN082 (10 mg/kg) groups. (B) EMG signal recordings of an NMS rat and an NMS + AMN082 (10 mg/kg) rat in response to CRD. (C) Statistical analysis of the EMG amplitude among the groups. (D) Statistical analysis of the AWR scores among the groups. AWR data were analysed by one-way ANOVA and least significant difference test; EMG data were analysed by one-way ANOVA and Dunnett's T3 test. Data are expressed as the means ± standard error of the mean (number of rats=6). * P<0.05, ** P<0.01. ANOVA, analysis of variance; AUC, area under the curve; AWR, abdominal withdrawal reflex; CRD, colorectal distension; EMG, electromyography; mGluR7, metabotropic glutamate receptor-7; NMS, neonatal maternal separation.

Article Snippet: Tissues were incubated overnight at 4°C with anti-mGluR7 antibody (1:500, rabbit anti-rat, cat. no. NB-91787; Novus Biologicals LLC), or antibodies against cluster of differentiation (CD)3 (1:100, rabbit anti-rat, cat. no. ab16669) CD68 (1:200, mouse anti-rat, cat. no. ab31630) or myeloperoxidase (MPO; 1:50, rabbit anti-rat, cat. no. ab9535) (Abcam, Cambridge, UK).

Techniques: Expressing

Journal: American Journal of Translational Research

Article Title: IL-4 and CCR7 play an important role in the development of keloids in patients with a family history

doi:

Figure Lengend Snippet: Primers and their sequences for PCR analysis

Article Snippet: CCR7 polyclonal antibody (dilution rate = 1:1000, 25898-1-AP, Proteintech, USA) was used to detect CCR7, while the secondary antibody was a goat anti-mouse monoclonal antibody (dilution rate = 1:5000, ab205719, Abcam, Cambridge, UK).

Techniques: Sequencing

Journal: American Journal of Translational Research

Article Title: IL-4 and CCR7 play an important role in the development of keloids in patients with a family history

doi:

Figure Lengend Snippet: A. The network of enriched terms colored by cluster ID, where nodes that share the same cluster ID are typically close to each other. B. The network of enriched terms colored by P-values, where terms containing more genes tend to have a more significant P-value. C-H. Relative expression of CCR7, CD40LG, IL-4, IL-10, CD80, and CTLA-4 by RT-qPCR analysis.

Article Snippet: CCR7 polyclonal antibody (dilution rate = 1:1000, 25898-1-AP, Proteintech, USA) was used to detect CCR7, while the secondary antibody was a goat anti-mouse monoclonal antibody (dilution rate = 1:5000, ab205719, Abcam, Cambridge, UK).

Techniques: Expressing, Quantitative RT-PCR

Journal: American Journal of Translational Research

Article Title: IL-4 and CCR7 play an important role in the development of keloids in patients with a family history

doi:

Figure Lengend Snippet: A-D. Relative expression of CD86, IL-2, IL-6, and STAT3 by RT-qPCR analysis. E. Western blotting analysis showed that the expression of IL-4 and CCR7 proteins between the FN and N groups. F. The expression of IL-4 between the FN and N groups was investigated using immunohistochemistry, × 20 and × 100.

Article Snippet: CCR7 polyclonal antibody (dilution rate = 1:1000, 25898-1-AP, Proteintech, USA) was used to detect CCR7, while the secondary antibody was a goat anti-mouse monoclonal antibody (dilution rate = 1:5000, ab205719, Abcam, Cambridge, UK).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Immunohistochemistry

Journal: American Journal of Translational Research

Article Title: IL-4 and CCR7 play an important role in the development of keloids in patients with a family history

doi:

Figure Lengend Snippet: A. The expression of CCR7 between the FN and N groups was investigated using immunohistochemistry, × 20 and × 100. B. The expression of IL-4 was investigated between the FN and N groups using immunofluorescence, × 100 and × 400 (enlarge). C. The expression of CCR7 between the FN and N groups was investigated using immunofluorescence, × 100 and × 400 (enlarge). D. The mean fluorescence intensity of IL-4 was analyzed by Image-Pro Plus 6.0. E. The mean fluorescence intensity of CCR7 was analyzed by Image-Pro Plus 6.0.

Article Snippet: CCR7 polyclonal antibody (dilution rate = 1:1000, 25898-1-AP, Proteintech, USA) was used to detect CCR7, while the secondary antibody was a goat anti-mouse monoclonal antibody (dilution rate = 1:5000, ab205719, Abcam, Cambridge, UK).

Techniques: Expressing, Immunohistochemistry, Immunofluorescence, Fluorescence

Journal: Molecular Pain

Article Title: Effects of a metabotropic glutamate receptor subtype 7 negative allosteric modulator in the periaqueductal grey on pain responses and rostral ventromedial medulla cell activity in rat

doi: 10.1186/1744-8069-9-44

Figure Lengend Snippet: Expression of mGluR7 mRNA in the VL PAG: RT-PCR analysis starting from 100ng mRNA shows an increase for mGluR7 gene expression in SNI rats compared to sham rats. Quantification of the expression levels is reported in the graphic that shows RT-PCR data value for mGluR7 relative to the housekeeping 18S gene and normalized with respect to the mean of the shams (A) . Western blot analysis for mGluR7 protein in PAG lysate from SNI and sham rats normalized with respect to β-tubulin and sham mean (B) . Expression of mGluR7 in the PAG of SNI and sham rats. Double staining shows the high expression of mGluR7 on vGluT positive terminals (right) and low expression on VGAT positive terminals (left). Wave length is 568 nm for red staining and 488 for green staining (not shown) the merge was performed of 488 and 568 channels (yellow). Lowest panel indicate the double staining mGluR7 (red) synaptophysin (green) and relative merge (C) . Purple spots indicate a DAPI and red background/fluorescence spot overlapping. Scale bar = 100 μm. Data are represented as a mean ± SEM. p < 0.05 was considered statistically significant.

Article Snippet: Sequences for rat mGluR7 mRNAs from GenBank (DNASTAR Inc.) were used to design primer pairs for RT-PCRs (OLIGO 4.05 software, National Biosciences Inc.).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Gene Expression, Western Blot, Double Staining, Staining, Fluorescence

Journal: PLoS ONE

Article Title: Ionizing Radiation Induces Altered Neuronal Differentiation by mGluR1 through PI3K-STAT3 Signaling in C17.2 Mouse Neural Stem-Like Cells

doi: 10.1371/journal.pone.0147538

Figure Lengend Snippet: Inhibitors or antagonists used in this study.

Article Snippet: For the experiments using inhibitors or antagonists on mitogen-activated protein kinase kinase (MEK) (PD98059; Cell Signaling Technologies, MA, USA), protein kinase A (PKA) (H-89; Cell Signaling Technologies), p38 (SB203580; Cell Signaling Technologies), PI3k (LY294002; Cell Signaling Technologies), tropomyosin receptor kinase A (Trk A) (AG879; Tocris, Bristol, UK), tropomyosin receptor kinase B (Trk B) (ANA12; Tocris), ataxia telangiectasia mutated (ATM) (KU55933; Selleckchem, TX, USA), ataxia telangiectasia and Rad3-related protein (ATR) (VE821; Selleckchem), c-Jun N-terminal kinase (JNK) (SP600125; Selleckchem), signal transducers and activators of transcription factors 1 (STAT1) (Fludarabine; Selleckchem), signal transducers and activators of transcription factors 3 (STAT3) (S3I-201; Selleckchem), GTPase (Rac) (NSC23766; Selleckchem), rapidly accelerated fibrosarcoma kinase (B-Raf) (GDC-0879; Selleckchem), metabotropic glutamate receptor 1 (mGluR1) (LY367385; Tocris), mGluR5 (MPEP; Tocris), Group II mGluRs (mGluR2 and mGluR3) (LY341495; Tocris) and Group III mGluRs (mGluR4, mGluR7 and mGluR8) (MSOP; Tocris), Neuro-2a cells were treated with these inhibitors at 5 or 10 μM and C17.2 cells were treated with theses inhibitors or antagonists at 0.1~25 μM, 2 hr prior to the irradiation.

Techniques:

Journal: PLoS ONE

Article Title: Ionizing Radiation Induces Altered Neuronal Differentiation by mGluR1 through PI3K-STAT3 Signaling in C17.2 Mouse Neural Stem-Like Cells

doi: 10.1371/journal.pone.0147538

Figure Lengend Snippet: Primer sequences for real-time PCR used in this study.

Article Snippet: For the experiments using inhibitors or antagonists on mitogen-activated protein kinase kinase (MEK) (PD98059; Cell Signaling Technologies, MA, USA), protein kinase A (PKA) (H-89; Cell Signaling Technologies), p38 (SB203580; Cell Signaling Technologies), PI3k (LY294002; Cell Signaling Technologies), tropomyosin receptor kinase A (Trk A) (AG879; Tocris, Bristol, UK), tropomyosin receptor kinase B (Trk B) (ANA12; Tocris), ataxia telangiectasia mutated (ATM) (KU55933; Selleckchem, TX, USA), ataxia telangiectasia and Rad3-related protein (ATR) (VE821; Selleckchem), c-Jun N-terminal kinase (JNK) (SP600125; Selleckchem), signal transducers and activators of transcription factors 1 (STAT1) (Fludarabine; Selleckchem), signal transducers and activators of transcription factors 3 (STAT3) (S3I-201; Selleckchem), GTPase (Rac) (NSC23766; Selleckchem), rapidly accelerated fibrosarcoma kinase (B-Raf) (GDC-0879; Selleckchem), metabotropic glutamate receptor 1 (mGluR1) (LY367385; Tocris), mGluR5 (MPEP; Tocris), Group II mGluRs (mGluR2 and mGluR3) (LY341495; Tocris) and Group III mGluRs (mGluR4, mGluR7 and mGluR8) (MSOP; Tocris), Neuro-2a cells were treated with these inhibitors at 5 or 10 μM and C17.2 cells were treated with theses inhibitors or antagonists at 0.1~25 μM, 2 hr prior to the irradiation.

Techniques: Real-time Polymerase Chain Reaction, Sequencing

Journal: PLoS ONE

Article Title: Ionizing Radiation Induces Altered Neuronal Differentiation by mGluR1 through PI3K-STAT3 Signaling in C17.2 Mouse Neural Stem-Like Cells

doi: 10.1371/journal.pone.0147538

Figure Lengend Snippet: Summary of expression profile of neuronal function-related genes in IR-stimulated and neurotrophin-stimulated C17.2 cells.

Article Snippet: For the experiments using inhibitors or antagonists on mitogen-activated protein kinase kinase (MEK) (PD98059; Cell Signaling Technologies, MA, USA), protein kinase A (PKA) (H-89; Cell Signaling Technologies), p38 (SB203580; Cell Signaling Technologies), PI3k (LY294002; Cell Signaling Technologies), tropomyosin receptor kinase A (Trk A) (AG879; Tocris, Bristol, UK), tropomyosin receptor kinase B (Trk B) (ANA12; Tocris), ataxia telangiectasia mutated (ATM) (KU55933; Selleckchem, TX, USA), ataxia telangiectasia and Rad3-related protein (ATR) (VE821; Selleckchem), c-Jun N-terminal kinase (JNK) (SP600125; Selleckchem), signal transducers and activators of transcription factors 1 (STAT1) (Fludarabine; Selleckchem), signal transducers and activators of transcription factors 3 (STAT3) (S3I-201; Selleckchem), GTPase (Rac) (NSC23766; Selleckchem), rapidly accelerated fibrosarcoma kinase (B-Raf) (GDC-0879; Selleckchem), metabotropic glutamate receptor 1 (mGluR1) (LY367385; Tocris), mGluR5 (MPEP; Tocris), Group II mGluRs (mGluR2 and mGluR3) (LY341495; Tocris) and Group III mGluRs (mGluR4, mGluR7 and mGluR8) (MSOP; Tocris), Neuro-2a cells were treated with these inhibitors at 5 or 10 μM and C17.2 cells were treated with theses inhibitors or antagonists at 0.1~25 μM, 2 hr prior to the irradiation.

Techniques: Expressing, Functional Assay, Control

Journal: PLoS ONE

Article Title: Ionizing Radiation Induces Altered Neuronal Differentiation by mGluR1 through PI3K-STAT3 Signaling in C17.2 Mouse Neural Stem-Like Cells

doi: 10.1371/journal.pone.0147538

Figure Lengend Snippet: C17.2 cells treated with the antagonists of mGluR1 (LY367385) at 25 μM, mGluR5 (MPEP) at 5 μM, Group II mGluRs (mGluR2 and mGluR3; LY341495) at 100 nM and Group III mGluRs (mGluR4, mGluR7 and mGluR8; MSOP) at 100 μM for 2 hr, and then exposed to IR at 6 Gy and incubated at 37°C for 72 hr. To assess rate of neurite bearing cells, each 200 cells in three randomly taken images were analyzed by Image J software. IR-induced neurite outgrowth is not affected by the antagonists of mGluR5, Group II mGluRs nad Group III mGluRs but was blocked by mGluR1 antagonist, LY367385 (A). Expression of NSC marker, nestin, and neuronal marker, β-III tubulin was analyzed by Western blot (B, upper panel), and then quantified by Image J software (B, down panel). To assess effect of mGluR1 inhibition on IR-induced neuronal function-related genes, mRNA expression of synaptophysin, synaptotagmin1 and GABA A receptors was analyzed by real-time PCR (C). *p < 0.05, **p < 0.01 vs naive group, †p < 0.05, ††p < 0.01 vs radiation only group.

Article Snippet: For the experiments using inhibitors or antagonists on mitogen-activated protein kinase kinase (MEK) (PD98059; Cell Signaling Technologies, MA, USA), protein kinase A (PKA) (H-89; Cell Signaling Technologies), p38 (SB203580; Cell Signaling Technologies), PI3k (LY294002; Cell Signaling Technologies), tropomyosin receptor kinase A (Trk A) (AG879; Tocris, Bristol, UK), tropomyosin receptor kinase B (Trk B) (ANA12; Tocris), ataxia telangiectasia mutated (ATM) (KU55933; Selleckchem, TX, USA), ataxia telangiectasia and Rad3-related protein (ATR) (VE821; Selleckchem), c-Jun N-terminal kinase (JNK) (SP600125; Selleckchem), signal transducers and activators of transcription factors 1 (STAT1) (Fludarabine; Selleckchem), signal transducers and activators of transcription factors 3 (STAT3) (S3I-201; Selleckchem), GTPase (Rac) (NSC23766; Selleckchem), rapidly accelerated fibrosarcoma kinase (B-Raf) (GDC-0879; Selleckchem), metabotropic glutamate receptor 1 (mGluR1) (LY367385; Tocris), mGluR5 (MPEP; Tocris), Group II mGluRs (mGluR2 and mGluR3) (LY341495; Tocris) and Group III mGluRs (mGluR4, mGluR7 and mGluR8) (MSOP; Tocris), Neuro-2a cells were treated with these inhibitors at 5 or 10 μM and C17.2 cells were treated with theses inhibitors or antagonists at 0.1~25 μM, 2 hr prior to the irradiation.

Techniques: Incubation, Software, Expressing, Marker, Western Blot, Inhibition, Real-time Polymerase Chain Reaction