Journal: The Journal of Headache and Pain

Article Title: PACAP6-38 improves nitroglycerin-induced central sensitization by modulating synaptic plasticity at the trigeminal nucleus caudalis in a male rat model of chronic migraine

doi: 10.1186/s10194-023-01603-3

Figure Lengend Snippet: Antibodies used in western blot and immunofluorescence staining assays

Article Snippet: SYT1 , Abclonal , A0992 , Rabbit , 1:1000.

Techniques: Western Blot, Immunofluorescence, Staining

Journal: Frontiers in Molecular Neuroscience

Article Title: Proteomic analysis of spinal cord tissue in a rat model of cancer-induced bone pain

doi: 10.3389/fnmol.2022.1009615

Figure Lengend Snippet: The detail information of the top 30 up-regulated differentially expressed proteins.

Article Snippet: The following antibodies from ABclonal Technology were used: CPLX1 rabbit pAb (1:1000, A11588), SNAP25 rabbit pAb (1:1000, A2234), SYT1 rabbit pAb (1:1000, A0992), ALDH1B1 rabbit pAb (1:1000, A3725), GATM rabbit pAb (1:1000, A6598), NDUFA11 rabbit pAb (1:1000, A16239), and HRP-conjugated secondary antibodies (1:5000, AS014).

Techniques:

Journal: Frontiers in Molecular Neuroscience

Article Title: Proteomic analysis of spinal cord tissue in a rat model of cancer-induced bone pain

doi: 10.3389/fnmol.2022.1009615

Figure Lengend Snippet: The information of up and down-regulated differentially expressed synaptic proteins.

Article Snippet: The following antibodies from ABclonal Technology were used: CPLX1 rabbit pAb (1:1000, A11588), SNAP25 rabbit pAb (1:1000, A2234), SYT1 rabbit pAb (1:1000, A0992), ALDH1B1 rabbit pAb (1:1000, A3725), GATM rabbit pAb (1:1000, A6598), NDUFA11 rabbit pAb (1:1000, A16239), and HRP-conjugated secondary antibodies (1:5000, AS014).

Techniques:

Journal: Frontiers in Molecular Neuroscience

Article Title: Proteomic analysis of spinal cord tissue in a rat model of cancer-induced bone pain

doi: 10.3389/fnmol.2022.1009615

Figure Lengend Snippet: Interaction networks analyzed for differentially expressed proteins (DEPs). (A) STRING protein–protein interaction network of DEPs colored nodes indicates the individual protein identified. Lines between nodes represent the direct and indirect association of proteins. Red color presented up-regulated proteins. Green color presented down-regulated proteins. Circle 1: synaptic interaction proteins such as CPLX1, SNAP25, and SYT1 proteins. Circle 2: mitochondrial interaction proteins such as ALDH1B and GATM. (B) Heatplot presents differentially expressed proteins related to the pathway. Red ovals indicated proteins such as CPLX1, SNAP25, and SYT1 involved in the synaptic vesicle cycle pathway, and ALDH1B and GATM involved in arginine and proline metabolism.

Article Snippet: The following antibodies from ABclonal Technology were used: CPLX1 rabbit pAb (1:1000, A11588), SNAP25 rabbit pAb (1:1000, A2234), SYT1 rabbit pAb (1:1000, A0992), ALDH1B1 rabbit pAb (1:1000, A3725), GATM rabbit pAb (1:1000, A6598), NDUFA11 rabbit pAb (1:1000, A16239), and HRP-conjugated secondary antibodies (1:5000, AS014).

Techniques:

Journal: Frontiers in Molecular Neuroscience

Article Title: Proteomic analysis of spinal cord tissue in a rat model of cancer-induced bone pain

doi: 10.3389/fnmol.2022.1009615

Figure Lengend Snippet: Validation of synaptic- and mitochondrial-related proteins using immunofluorescence and Western blot assay. (A,B) Representative immunofluorescence staining images (A) and quantitative analysis (B) for SNAP25, GATM, and NDUFA11 in the spinal cord. Scale bar = 20 μm. Data were expressed as the mean ± SD ( n = 3 mice/group). * p < 0.05 vs. sham group. (C,D) Representative Western blot bands (C) and quantitative analysis (D) of CPLX1, SNAP25, and SYT1 protein in the spinal cord. Data were presented as mean ± SD ( n = 3 mice/group). * p < 0.05 vs. sham group. (E,F) Representative western blot bands (E) and quantitative analysis (F) of ALDH1B, GATM, and NDUFA11 protein in spinal cord. Data were presented as mean ± SD ( n = 3 mice/group). * p < 0.05 vs. sham group.

Article Snippet: The following antibodies from ABclonal Technology were used: CPLX1 rabbit pAb (1:1000, A11588), SNAP25 rabbit pAb (1:1000, A2234), SYT1 rabbit pAb (1:1000, A0992), ALDH1B1 rabbit pAb (1:1000, A3725), GATM rabbit pAb (1:1000, A6598), NDUFA11 rabbit pAb (1:1000, A16239), and HRP-conjugated secondary antibodies (1:5000, AS014).

Techniques: Biomarker Discovery, Immunofluorescence, Western Blot, Staining

Journal: Journal of Orthopaedic Translation

Article Title: Metformin improves fibroblast metabolism and ameliorates arthrofibrosis in rats

doi: 10.1016/j.jot.2023.05.011

Figure Lengend Snippet: Glycolysis is upregulated in human fibrotic capsules. a The capsule tissue samples were obtained from patients with or without shoulder adhesive capsulitis during arthroscopic shoulder surgery and were stained with HE and Masson's trichrome (n ​= ​8, black arrows: densely packed collagen fibers and fibroblastic proliferation, scale bars: upper panel: 200 ​μm, lower panel: 20 ​μm). Harvested tissues were homogenized, and mRNA ( b ) or protein ( c-d ) levels of α-SMA, FN1, TGF-β, GLUT1, HK2, PFKFB3, PFKP, PKM, and LDHA were quantified by qRT-PCR (n ​= ​8) or western blotting (n ​= ​5), respectively. The data were normalized to β-actin and expressed as value relative to the level in normal capsular tissue that was designated 1. e-f The capsular tissue sections were subjected to IHC staining for α-SMA, GLUT1, and LDHA, and the positive area was quantified (n ​= ​5, scale bars: upper panel: 200 ​μm, lower panel: 10 ​μm). The data are shown as mean ​± ​SEM. ∗ P <0.05, ∗∗ P <0.01, ∗∗∗ P <0.001, ∗∗∗∗ P <0.0001 by unpaired Student's t test.

Article Snippet: Antibodies against Glucose transporter 1 (GLUT1) (ABclonal, A11727), Hexokinase-2 (HK2) (ABclonal, A0994), Phosphofructokinase (PFKP) (ABclonal, A12160), Pyruvate kinase M−2 (PKM2) (ABclonal, A16700), α-SMA (ABclonal, A1011), Fibronectin-1 (FN1) (ABclonal, A7488), LDHA (ABclonal, A0861), and β-actin (Zen Bio, 200,068-8F10) were used.

Techniques: Capsules, Adhesive, Staining, Quantitative RT-PCR, Western Blot, Immunohistochemistry

Journal: Journal of Orthopaedic Translation

Article Title: Metformin improves fibroblast metabolism and ameliorates arthrofibrosis in rats

doi: 10.1016/j.jot.2023.05.011

Figure Lengend Snippet: Metformin inhibits fibroblast activation and glycolytic capacity. Primary human fibroblasts were treated with PBS (control), TGF-β (10 ​ng/ml), or TGF-β (10 ​ng/ml) ​+ ​metformin (2 ​mM) for 48 ​h. a-b Immunofluorescent staining of α-SMA and FN1. c-d The results of quantitative analysis showed a significant decrease in α-SMA and FN1 expression in fibroblasts treated with metformin (n ​= ​3). e The effects of metformin on the protein synthesis of α-SMA, FN1, GLUT1, HK2, PFKP, PKM2, and LDHA. f Statistical analysis of protein expression levels shown in (e) (n ​= ​3). Protein abundance was normalized to β-actin and expressed as relative value to the level in control cells that was designated 1. g mRNA levels of α-SMA, FN1, GLUT1, HK2, PFKP, PKM, and LDHA were quantified by qRT-PCR (n ​= ​3). Data was normalized to β-actin and expressed as value relative to the level in control cells that was designated 1. The data are shown as mean ​± ​SEM. ∗comparison between control and TGF-β treated cells, # comparison between TGF-β and TGF-β ​+ ​metformin treated cells. ∗ P <0.05, ∗∗ P <0.01, ∗∗∗ P <0.001, ∗∗∗∗ P <0.0001 by one-way ANOVA followed by Tukey–Kramer test. Scale bars: 50 ​μm.

Article Snippet: Antibodies against Glucose transporter 1 (GLUT1) (ABclonal, A11727), Hexokinase-2 (HK2) (ABclonal, A0994), Phosphofructokinase (PFKP) (ABclonal, A12160), Pyruvate kinase M−2 (PKM2) (ABclonal, A16700), α-SMA (ABclonal, A1011), Fibronectin-1 (FN1) (ABclonal, A7488), LDHA (ABclonal, A0861), and β-actin (Zen Bio, 200,068-8F10) were used.

Techniques: Activation Assay, Control, Staining, Expressing, Quantitative Proteomics, Quantitative RT-PCR, Comparison

Journal: Cancer Medicine

Article Title: Construction of a prognostic glycolysis‐related lncRNA signature for patients with colorectal cancer

doi: 10.1002/cam4.4851

Figure Lengend Snippet: In vitro experiments. (A–B) High expression of AFAP1‐AS1 indicates worse OS in both TCGA and GSE39582 datasets. (C) Efficiency of si‐AFAP1‐AS1 was detected by RT‐qPCR. (D–E) Glucose uptake assay and lactate production assay in DLD1 cells that were transfected with siAFAP1‐AS1 or siNC. (F) Protein level of HK2, PKM2, LDHA after knocking down AFAP1‐AS1. (G) Effects of AFAP1‐AS1 on the invasion ability of DLD1 cells were measured by transwell assay. (H) Effects of AFAP1‐AS1 on the migration ability of DLD1 cells were detected by wound healing assay.

Article Snippet: Proteins were probed using primary antibodies against HK2 (A0994, 1:1000; ABclonal), LDHA (A1146, 1:1000; ABclonal), PKM2 (A19102, 1:1000; ABclonal), β‐actin (ab133626, 1:5000; Abcam).

Techniques: In Vitro, Expressing, Quantitative RT-PCR, Transfection, Transwell Assay, Migration, Wound Healing Assay