Journal: Brain Structure & Function

Article Title: Altered motor, anxiety-related and attentional task performance at baseline associate with multiple gene copies of the vesicular acetylcholine transporter and related protein overexpression in ChAT::Cre+ rats

doi: 10.1007/s00429-019-01957-y

Figure Lengend Snippet: Measurement of prefrontal cortical VAChT mRNA and protein levels. a qPCR results indicated additional mouse VAChT gene copies in all ChAT::Cre+ rats ( n = 5) compared to ChAT::Cre− controls ( n = 8). This translated to b increased mouse VAChT mRNA expression in Cre+ ( n = 5) and Cre− ( n = 7) siblings compared to Wt mice ( n = 4) and also c increased VAChT protein expression in PFC brain samples of Cre+ ( n = 5) and Cre− ( n = 8) rats. All data are presented as the mean ± SEM. * p ≤ 0.05, ** p ≤ 0.01 and *** p < 0.001, compared to ChAT::Cre− rats and Wt mouse

Article Snippet: Proteins were transferred to Immobilon-P polyvinylidene difluoride (PVDF) membranes (Cat No. IPVH00010, Millipore, UK) using a semi-dry transfer apparatus (Trans-Blot TurboTM, Bio-Rad) run at 20 V for 1 h. The membrane was then blocked in 5% (w/v) powdered skimmed milk in Tris-buffered saline/0.1% Tween 20 (TBS-T) for 1 h, followed by incubation overnight at 4 °C with a primary antibody raised against α-VAChT (1:1000, rabbit polyclonal, Cat No. 139-103, Synaptic Systems, Göttingen, Germany; RRID: AB_10893979) diluted in blocking solution.

Techniques: Expressing

Journal: Scientific Reports

Article Title: Viral-like TLR3 induction of cytokine networks and α-synuclein are reduced by complement C3 blockade in mouse brain

doi: 10.1038/s41598-023-41240-z

Figure Lengend Snippet: Intra-striatal TLR3 activation in mouse brain increased complement C3 and α-synuclein protein levels. WT mice received a single unilateral deposit of 50 μg Poly(I:C) (n = 10) or vehicle (n = 9) in the striatum. Two days later, injected striata were isolated and homogenised in RIPA buffer for protein detection by Western blot ( A, E ). ( B – D, F – I ) Histogram bars represent the mean levels of target protein normalized to GAPDH or total protein visualized by stain-free UV-fluorescence. Uncropped blots can be found in Figs. – . Error bars represent SEM. A.U., arbitrary units. Statistical analysis was performed using unpaired t-tests. ns, p > 0.05; *, p < 0.05; ***, p < 0.001.

Article Snippet: Membranes were blocked for 30 min with 5% milk in Tris-buffered saline containing 0.1% Tween 20 and were probed overnight at 4 °C with primary antibodies against the following target proteins: α-synuclein (BD biosciences, BD610787 1:2000), C3 (Abcam, ab200999, 1:1000), C1q (Abcam, ab182451, 1:500), Iba1 (Abcam, ab178846, 1:1000), GFAP (EMD Millipore, MAB360, 1:2000), GAPDH (Cell Signalling Technology, #2118, 1:2000).

Techniques: Activation Assay, Injection, Isolation, Western Blot, Staining, Fluorescence

Journal: Scientific Reports

Article Title: Viral-like TLR3 induction of cytokine networks and α-synuclein are reduced by complement C3 blockade in mouse brain

doi: 10.1038/s41598-023-41240-z

Figure Lengend Snippet: Knockdown of C3 prior to TLR3 activation reduced α-synuclein protein levels in the striatum of mouse brain. Alpha-synuclein levels in the injected striatum of mice from NT ASO/Poly(I:C) (n = 10) and C3 ASO/Poly(I:C) (n = 10) groups were measured by Western blot ( A ). The uncropped blot can be found in Fig. . ( B ) Histogram bars represent the mean levels of α-synuclein normalized to total protein loading. Error bars represent SEM. A.U., arbitrary units. Statistical analysis was performed using unpaired t-tests. ***, p < 0.001.

Article Snippet: Membranes were blocked for 30 min with 5% milk in Tris-buffered saline containing 0.1% Tween 20 and were probed overnight at 4 °C with primary antibodies against the following target proteins: α-synuclein (BD biosciences, BD610787 1:2000), C3 (Abcam, ab200999, 1:1000), C1q (Abcam, ab182451, 1:500), Iba1 (Abcam, ab178846, 1:1000), GFAP (EMD Millipore, MAB360, 1:2000), GAPDH (Cell Signalling Technology, #2118, 1:2000).

Techniques: Activation Assay, Injection, Western Blot

Journal: International Orthopaedics

Article Title: Abnormal expression of chondroitin sulphate N-acetylgalactosaminyltransferase 1 and Hapln-1 in cartilage with Kashin–Beck disease and primary osteoarthritis

doi: 10.1007/s00264-013-1937-y

Figure Lengend Snippet: Chondrocyte gene network of CSGalNAcT-1 was classified as glycan biosynthesis and metabolism and functional pathway

Article Snippet: For the primary antibody, polyclonal rabbit anti-human primary antibodies against CSGalNAcT-1 (1:50; ABGENT, San Diego, CA 92121, USA) and Hapln-1 (1:50; Abcam, UK) were used at 4 ° C overnight.

Techniques: Functional Assay

Journal: International Orthopaedics

Article Title: Abnormal expression of chondroitin sulphate N-acetylgalactosaminyltransferase 1 and Hapln-1 in cartilage with Kashin–Beck disease and primary osteoarthritis

doi: 10.1007/s00264-013-1937-y

Figure Lengend Snippet: CSGalNAcT-1 and Hapln-1 mRNA expression in osteoarthritis (OA) cartilage, Kashin-Beck disease (KBD) cartilage and normal cartilage. CSGalNAcT-1 and Hapln-1 mRNA levels reach statistical significance. * Denotes significant difference at P < 0.05

Article Snippet: For the primary antibody, polyclonal rabbit anti-human primary antibodies against CSGalNAcT-1 (1:50; ABGENT, San Diego, CA 92121, USA) and Hapln-1 (1:50; Abcam, UK) were used at 4 ° C overnight.

Techniques: Expressing

Journal: International Orthopaedics

Article Title: Abnormal expression of chondroitin sulphate N-acetylgalactosaminyltransferase 1 and Hapln-1 in cartilage with Kashin–Beck disease and primary osteoarthritis

doi: 10.1007/s00264-013-1937-y

Figure Lengend Snippet: Comparison of CSGalNAcT-1 expression in the cartilage from osteoarthritis (OA), Kashin-Beck disease (KBD) and normal groups. a , b , c Denote the upper of normal, OA and KBD cartilage, respectively. d , e , f Denote the middle of normal, OA and KBD cartilage, respectively. g , h , i Denote the deep of normal, OA and KBD cartilage, respectively

Article Snippet: For the primary antibody, polyclonal rabbit anti-human primary antibodies against CSGalNAcT-1 (1:50; ABGENT, San Diego, CA 92121, USA) and Hapln-1 (1:50; Abcam, UK) were used at 4 ° C overnight.

Techniques: Expressing

Journal: International Orthopaedics

Article Title: Abnormal expression of chondroitin sulphate N-acetylgalactosaminyltransferase 1 and Hapln-1 in cartilage with Kashin–Beck disease and primary osteoarthritis

doi: 10.1007/s00264-013-1937-y

Figure Lengend Snippet: CSGalNAcT-1 and Hapln-1 protein level expression reduced in osteoarthritis (OA) and Kashin-Beck disease (KBD) cartilage compared with normal controls. Wnt 3a, β-catenin and Runx-2 level expression increased in OA and KBD cartilage compared with normal controls. A KBD free-Se group. B 0.25 μg/ml Se + KBD group. C 0.10 μg/ml Se + KBD group. D 0.05 μg/ml Se + KBD group

Article Snippet: For the primary antibody, polyclonal rabbit anti-human primary antibodies against CSGalNAcT-1 (1:50; ABGENT, San Diego, CA 92121, USA) and Hapln-1 (1:50; Abcam, UK) were used at 4 ° C overnight.

Techniques: Expressing

Journal: PLoS ONE

Article Title: Glycoprotein Nonmetastatic Melanoma B (Gpnmb)-Positive Macrophages Contribute to the Balance between Fibrosis and Fibrolysis during the Repair of Acute Liver Injury in Mice

doi: 10.1371/journal.pone.0143413

Figure Lengend Snippet: (A) Hepatic expression of IL-1β, IL-10, and CCL2 are significantly decreased (clodronate liposome injection group [ n = 3, only survival] vs. PBS injection group [ n = 4)]). Values are mean ± SEM. * P < 0.05 (Mann-Whitney U test)). (B) Collagen deposition and α-SMA expression are decreased (original magnification, x400). (C) Hepatic expression of TGF-β, Col1α1, and MMP-13 are significantly decreased (clodronate liposome injection group [ n = 3, only survival] vs. PBS injection group [ n = 4]). Values are mean ± SEM. * P < 0.05 (Mann-Whitney U test)).

Article Snippet: For immunohistochemical analysis, liver specimens were fixed in 10% buffered formalin or 4% paraformaldehyde and incubated with rat anti-mouse F4/80 (Clone: A3-1, AbD Serotec, Raleigh, NC, USA), rat anti-mouse CD68 (Clone: FA-11, AbD Serotec), goat anti-mouse Gpnmb (Clone: #297310, R&D Systems, Minneapolis, MN, USA), and monoclonal antibody against α-SMA (Clone: E184, Merck Millipore, Billerica, MA, USA).

Techniques: Expressing, Injection, MANN-WHITNEY

Journal: PLoS ONE

Article Title: Glycoprotein Nonmetastatic Melanoma B (Gpnmb)-Positive Macrophages Contribute to the Balance between Fibrosis and Fibrolysis during the Repair of Acute Liver Injury in Mice

doi: 10.1371/journal.pone.0143413

Figure Lengend Snippet: (A) Expression of Gpnmb is observed in DBA-g+ but not DBA mice following a single injection of CCl 4 . However, (B) sequential changes in serum ALT levels and the degree of liver injury are not affected by the lack of Gpnmb-positive macrophages (original magnification, x100). Conversely, (C) the areas of fibrosis and (D) the number of α-SMA-positive cells are significantly decreased in the liver tissues of DBA mice compared to DBA-g+ mice (original magnification, x200). Additionally, (E) although lack of Gpnmb-positive macrophages does not affect expression of TGF-β or Col1α1, expression of MMP-9, MMP-13, and TIMP-1 are significantly decreased in mice lacking Gpnmb expression at six or eight days after single injection of CCl 4 . Values are mean ± SEM ( n = 4). * P < 0.05 (Mann-Whitney U test).

Article Snippet: For immunohistochemical analysis, liver specimens were fixed in 10% buffered formalin or 4% paraformaldehyde and incubated with rat anti-mouse F4/80 (Clone: A3-1, AbD Serotec, Raleigh, NC, USA), rat anti-mouse CD68 (Clone: FA-11, AbD Serotec), goat anti-mouse Gpnmb (Clone: #297310, R&D Systems, Minneapolis, MN, USA), and monoclonal antibody against α-SMA (Clone: E184, Merck Millipore, Billerica, MA, USA).

Techniques: Expressing, Injection, MANN-WHITNEY

Journal: PPAR Research

Article Title: The Antifibrosis Effects of Peroxisome Proliferator-Activated Receptor δ on Rat Corneal Wound Healing after Excimer Laser Keratectomy

doi: 10.1155/2014/464935

Figure Lengend Snippet: Representative immunofluorescent staining images for α -SMA (a) two weeks after laser treatment and fibronectin (b) and collagen type III (c) four weeks after laser treatment. DAPI-stained nuclei are shown in blue, α -SMA is shown in green, and fibronectin and collagen type III are shown in red. Scale bars, 50 μ m. (d) mRNA expression of α -SMA, fibronectin, collagen type III, and collagen type I at four weeks is depicted in the bar graph. Data are presented as the mean ± SD ( n = 3-4), and significant differences were analyzed by ANOVA. * P < 0.05, ** P < 0.01, *** P < 0.001 versus PBS group. # P < 0.05, ## P < 0.01, ### P < 0.001 versus GW501516 group.

Article Snippet: The primary antibodies used were peptide-affinity purified goat antibody against α -SMA (1 : 100 dilution; Abgent, USA), mouse monoclonal antibody against Col3a1 (1 : 100 dilution; Santa Cruz, CA, USA), and mouse monoclonal antibody against fibronectin (1 : 100 dilution; Santa Cruz, CA, USA).

Techniques: Staining, Expressing