Journal: Diagnostics

Article Title: Salivary Biomarker Profile in Periodontal Diseases: A Cross-Sectional Study on Leptin, Adiponectin, and Calprotectin

doi: 10.3390/diagnostics15222822

Figure Lengend Snippet: Comparison of receiver operating characteristic (ROC) curves of salivary leptin, adiponectin, and calprotectin in distinguishing different periodontal conditions. ( a ) Comparison of ROC curves of salivary biomarkers for differentiating healthy and gingivitis groups. ( b ) Comparison of ROC curves of salivary biomarkers for differentiating healthy and periodontitis groups. ( c ) Comparison of ROC curves of salivary biomarkers for distinguishing the gingivitis and periodontitis groups.

Article Snippet: Salivary adiponectin concentrations were examined using a human adiponectin ELISA kit (Cat# E4685Hu, BT-laboratory, Shanghai, China) according to the manufacturer’s instructions.

Techniques: Comparison

Journal: bioRxiv

Article Title: Trim39 regulates neuronal apoptosis by acting as a SUMO-targeted E3 ubiquitin-ligase for the transcription factor NFATc3

doi: 10.1101/2020.09.29.317958

Figure Lengend Snippet: A. BHK cells were transfected with Trim17-GFP together with Flag-Trim39 or empty plasmid (as a negative control) for 24 h. Cells were then treated with 20 μM MG-132 for 5 h. The cells were subsequently harvested and lysates were subjected to immunoprecipitation using anti-Flag agarose beads (left panel) or GFP-Trap beads (right panel). Immunoprecipitates and total lysates were analyzed by western blot using anti-GFP and anti-Flag antibodies. B. Neuro2A cells were transfected with HA-NFATc3 together with Flag-Trim39 or empty plasmid for 24 h. Cells were then treated as in A and lysates were subjected to immunoprecipitation using anti-Flag (left panel) or anti-HA (right panel) antibodies. Immunoprecipitates and total lysates were analyzed by western blot using anti-HA and anti-Flag antibodies. C. Neuro2A cells were treated with 10 μM MG-132 for 4 h and then fixed and subjected to in situ PLA using rabbit anti-NFATc3 and mouse anti-Trim39 antibodies. Each bright red spot indicates that the two proteins are in close proximity. Negative control was obtained by omitting anti-Trim39 antibody. When indicated, cells were previously transfected with Trim39 for 24 h (overexpressed). Images were analyzed by confocal microscopy. To better visualize the differences in PLA intensity, maximum intensity projection was applied to the z-stacks of images on the left panel. To better determine the subcellular location of the NFATc3/Trim39 interaction, a single slice of the z-stack is presented on the right panel (endo 1 slice). Nuclear staining was performed using DAPI. D. Neuro2A cells were transduced with lentiviral particles expressing a control shRNA (sheGFP) or a specific shRNA against Trim39 (shTrim39#1) for 24 h. Transduced cells were selected using puromycin for two additional days and plated onto coverslips. The day after plating, cells were analyzed by immunofluorescence using two different antibodies against Trim39; in red: antibody from Origene, in green: antibody from Proteintech. Images were set to the same minimum and maximum intensity to allow signal intensity comparison. E. Additional coverslips from the experiment presented in D were treated as in C and z-stacks of images were subjected to maximum intensity projection.

Article Snippet: Rabbit polyclonal antibody against Trim39 was from Proteintech (#12757-1-AP).

Techniques: Transfection, Plasmid Preparation, Negative Control, Immunoprecipitation, Western Blot, In Situ, Confocal Microscopy, Staining, Transduction, Expressing, Control, shRNA, Immunofluorescence, Comparison

Journal: bioRxiv

Article Title: Trim39 regulates neuronal apoptosis by acting as a SUMO-targeted E3 ubiquitin-ligase for the transcription factor NFATc3

doi: 10.1101/2020.09.29.317958

Figure Lengend Snippet: A. Neuro2A cells were transfected with HA-NFATc3 together with empty plasmid or His-tagged ubiquitin, in the presence or absence of Flag-Trim39 or the inactive mutant Flag-Trim39-ΔRING for 24 h. Cells were then incubated with 20 μM MG-132 for 6 h before harvesting. The ubiquitinated proteins were purified using nickel beads and analyzed by western blotting using anti-HA antibody to detect ubiquitin-conjugated HA-NFATc3. In a separate SDS-PAGE, samples of the input lysates used for the purification were analyzed with antibodies against HA and Flag. B. Neuro2A cells were transduced with lentiviruses expressing a control shRNA (directed against eGFP) or three different shRNAs targeting Trim39. Following 24 h transduction and 48 h selection of transduced cells using puromycin, cells were plated in new dishes. Then, cells were transfected with HA-NFATc3 or His-tagged ubiquitin or both for 24 h, and treated as in A. In conditions indicated by a star (*), some material was lost during TCA precipitation of the input lysate without affecting the amount of proteins in the nickel bead purification. These data are representative of 4 independent experiments. C. In vitro translated HA-NFATc3 was first immunopurified from wheat germ extract using anti-HA antibody. Then, beads used for immunopurification of NFATc3 were incubated for 1 h at 37°C in the in vitro ubiquitination reaction mix (containing E1 and E2 enzymes) with purified recombinant GST-Trim39 (WT) or its inactive mutant GST-Trim39-C49S/C52S (mut) in the presence or the absence of ubiquitin as indicated. Poly-ubiquitinated forms of NFATc3 were detected by immunoblotting using an anti-NFATc3 antibody. The same membrane was immunoblotted with an anti-TRIM39 antibody to verify that similar amounts of recombinant WT GST-Trim39 and GST-Trim39-C49S/C52S were used in the assay. Note that in the presence of ubiquitin the unmodified form of WT GST-Trim39 is lower due to high Trim39 ubiquitination.

Article Snippet: Rabbit polyclonal antibody against Trim39 was from Proteintech (#12757-1-AP).

Techniques: Transfection, Plasmid Preparation, Ubiquitin Proteomics, Mutagenesis, Incubation, Purification, Western Blot, SDS Page, Transduction, Expressing, Control, shRNA, Selection, TCA Precipitation, In Vitro, Immu-Puri, Recombinant, Membrane

Journal: bioRxiv

Article Title: Trim39 regulates neuronal apoptosis by acting as a SUMO-targeted E3 ubiquitin-ligase for the transcription factor NFATc3

doi: 10.1101/2020.09.29.317958

Figure Lengend Snippet: A. BHK cells were transfected with a fixed amount of a HA-NFATc3 expressing vector (1 μg) together with empty plasmid (−) or increasing amounts of Flag-Trim39 expressing vector (0.1, 0.2, 0.5 and 1 μg) or 0.2 μg of a vector expressing the inactive mutant Flag-Trim39-ΔRING. When indicated, the cells were treated with 10 μM MG-132 for 6 h before harvesting. Total lysates were analyzed by western blot using antibodies against HA, Flag and actin. B. Neuro2A cells were left untreated (NT), or transfected twice with an siRNA targeting specifically Trim39 (siTrim39#1) or with a negative control siRNA (siCtrl) for 48 h. Total lysates were analyzed by western blot using antibodies against NFATc3, Trim39 and actin. The intensity of the NFATc3 bands on the western blots was quantified, normalized by the intensity of the actin bands and expressed relative to the values obtained with the control shRNA. The graph shows mean ± SEM from three independent experiments. **P<0.01 significantly different from siCtrl (one-way ANOVA followed by Dunnett’s multiple comparisons test). C. Neuro2A cells were co-transfected with empty plasmid or HA-NFATc3, together with empty plasmid, Flag-Trim39 or the inactive mutant Flag-Trim39-ΔRING for 24 h. Then, total RNA was extracted and the mRNA level of Trim17 was estimated by quantitative PCR. Data are the means ± SEM of four independent experiments. **P<0.01 significantly different from the corresponding control (two-way ANOVA followed by Sidak’s multiple comparisons test). D. Neuro2A cells were transfected with empty plasmid, Flag-Trim39 or Flag-Trim39-ΔRING for 24 h. Then, cells were left untreated (control) or were deprived of serum for 3 h and subsequently treated with 1 μM A23187 and 100 nM PMA in serum-free medium for 1 h (A23+PMA). Total RNA was extracted and the mRNA level of Trim17 was estimated by quantitative PCR. Data are the means ± SEM of three independent experiments. *P<0.05; **P<0.01 significantly different from the corresponding value in cells transfected with empty plasmid (two-way ANOVA followed by Sidak’s multiple comparisons test). E. Neuro2A cells were transfected twice with two different siRNAs targeting specifically Trim39 or with a negative control siRNA for 48 h. Then, cells were left untreated (control) or were deprived of serum for 3h and subsequently treated with 1 μM A23187 and 100 nM PMA in serum-free medium for 30 min (A23+PMA). Total RNA was extracted and the mRNA level of Trim39 (left panel) or Trim17 (right panel) was estimated by quantitative PCR (NT = non transfected). Data are the means ± SEM of six independent experiments. *P<0.05; ***P<0.001 significantly different from cells transfected with control siRNA in the same condition (two-way ANOVA followed by Sidak’s multiple comparisons test).

Article Snippet: Rabbit polyclonal antibody against Trim39 was from Proteintech (#12757-1-AP).

Techniques: Transfection, Expressing, Plasmid Preparation, Mutagenesis, Western Blot, Negative Control, Control, shRNA, Real-time Polymerase Chain Reaction

Journal: bioRxiv

Article Title: Trim39 regulates neuronal apoptosis by acting as a SUMO-targeted E3 ubiquitin-ligase for the transcription factor NFATc3

doi: 10.1101/2020.09.29.317958

Figure Lengend Snippet: A. BHK cells were transfected with HA-NFATc3 together with His-tagged ubiquitin, in the presence or the absence of Flag-Trim39, Trim17-GFP or both, as indicated, for 24 h. Then, cells were incubated with 20 μM MG-132 for 6 h before harvesting. The ubiquitinated proteins were purified using nickel beads and analyzed by western blotting using anti-HA and anti-Flag antibodies to detect poly-ubiquitinated forms of NFATc3 and Trim39. In a separate SDS-PAGE, samples of the input lysates used for the purification were analyzed with antibodies against HA, Flag and GFP. B. In vitro translated HA-NFATc3 was first immunopurified from wheat germ extract using anti-HA antibody. Then, beads used for immunopurification of NFATc3 were incubated for 1 h at 37°C in the in vitro ubiquitination reaction mix (containing ubiquitin and E1 and E2 enzymes) with purified recombinant His-TRIM39 or MBP-TRIM17 as indicated. Poly-ubiquitinated forms of NFATc3, TRIM39 and TRIM17 were detected by immunoblotting using anti-NFATc3, anti-TRIM39 and anti-TRIM17 antibodies revealed using high exposure times. Low exposure times were used to compare the level of TRIM39 and TRIM17 in the different conditions.

Article Snippet: Rabbit polyclonal antibody against Trim39 was from Proteintech (#12757-1-AP).

Techniques: Transfection, Ubiquitin Proteomics, Incubation, Purification, Western Blot, SDS Page, In Vitro, Immu-Puri, Recombinant

Journal: bioRxiv

Article Title: Trim39 regulates neuronal apoptosis by acting as a SUMO-targeted E3 ubiquitin-ligase for the transcription factor NFATc3

doi: 10.1101/2020.09.29.317958

Figure Lengend Snippet: A,B. Neuro2A cells were transfected with HA-NFATc3 in the presence or the absence of Flag-Trim39, Trim17-GFP or both, as indicated, for 24 h. Cells were then treated with 20 μM MG-132 for 7 h. The cells were subsequently harvested and lysates were subjected to immunoprecipitation using anti-HA (A) or anti-Flag (B) antibodies. Immunoprecipitates and total lysates were analyzed by western blot using anti-HA, anti-GFP and anti-Flag antibodies. The intensity of the bands containing Flag-Trim39 co-immunoprecipitated with HA-NFATc3 was normalized by the intensity of the bands corresponding to immunoprecipitated HA-NFATc3 (A). The intensity of the bands containing HA-NFATc3 co-immunoprecipitated with Flag-Trim39 was normalized by the intensity of the bands corresponding to immunoprecipitated Flag-Trim39 (B). Relative values are indicated in red. C. Neuro2A cells were transfected with GFP or Trim17-GFP for 24 h. Then cells were treated with 10 μM MG-132 for 4 h, fixed and subjected to in situ PLA using rabbit anti-NFATc3 and mouse anti-Trim39 antibodies. Each bright red spot indicates that the two proteins are in close proximity. Images were analyzed by confocal microscopy and a single slice of the z-stacks is presented for each condition. Nuclear staining was performed using DAPI. Note that, in the Trim17-GFP condition, transfected cells (delineated by a yellow line) show less dots than neighboring non transfected cells, which is not the case in the GFP condition. D. The number of dots was determined in individual cells transfected with either GFP or Trim17-GFP using Fiji. Data represent one experiment, including 68 transfected cells for each condition, representative of two independent experiments. ****p<0.0001, significantly different from GFP transfected cells (unpaired t test).

Article Snippet: Rabbit polyclonal antibody against Trim39 was from Proteintech (#12757-1-AP).

Techniques: Transfection, Immunoprecipitation, Western Blot, In Situ, Confocal Microscopy, Staining

Journal: bioRxiv

Article Title: Trim39 regulates neuronal apoptosis by acting as a SUMO-targeted E3 ubiquitin-ligase for the transcription factor NFATc3

doi: 10.1101/2020.09.29.317958

Figure Lengend Snippet: A. Neuro2A cells were transfected with His-tagged ubiquitin together with WT HA-NFATc3 or HA-NFATc3 EallA, in the presence or the absence of Flag-Trim39, for 24 h. Then, cells were incubated with 20 μM MG-132 for 6 h before harvesting. The ubiquitinated proteins were purified using nickel beads and analyzed by western blotting using anti-HA antibody to detect ubiquitin-conjugated HA-NFATc3. In a separate SDS-PAGE, samples of the input lysates used for the purification were analyzed with antibodies against HA and Flag. B. Neuro2A cells were transfected with Flag-Trim39 together with WT HA-NFATc3, HA-NFATc3-EallA or empty plasmid for 24 h. Cells were then treated with 10 μM MG-132 for 8 h. The cells were subsequently harvested and lysates were subjected to immunoprecipitation using anti-HA antibody (left panel) or anti-Flag beads (right panel). Immunoprecipitates and total lysates were analyzed by western blot using anti-HA and anti-Flag antibodies. The intensity of the bands containing Flag-Trim39 co-immunoprecipitated with HA-NFATc3 was normalized by the intensity of the bands of immunoprecipitated HA-NFATc3. The intensity of the bands containing HA-NFATc3 co-immunoprecipitated with Flag-Trim39 was normalized by the intensity of the bands of immunoprecipitated Flag-Trim39. Relative values are indicated in red. C. Recombinant GST, GST-Trim39 and its different SIM mutants were purified using glutathione beads and subsequently incubated with purified recombinant SUMO-2 and SUMO-2 chains. Material bound to the beads was eluted and analyzed by western blot using anti-SUMO and anti-GST antibodies. A small fraction of the SUMO-2 chains was also loaded on the gel (input) for comparison. The intensity of bound SUMO-chain bands was quantified and normalized by the intensity of corresponding GST-Trim39 bands. Relative values are indicated in red. Note that SUMO bands are multiple of ≈ 15 kDa corresponding to mono-, di-, tri-, tetra-SUMO etc… D. Neuro2A cells were transfected with HA-NFATc3 or empty plasmid together with WT Flag-Trim39 or its SIM3 mutant for 24 h. Cells were treated as in B and lysates were subjected to immunoprecipitation using anti-HA antibody. Immunoprecipitates and total lysates were analyzed as in B. The intensity of the bands containing Flag-Trim39 co-immunoprecipitated with HA-NFATc3 was normalized by the intensity of the bands of immunoprecipitated HA-NFATc3. Relative values are indicated in red. E. Neuro2A cells were transfected with His-tagged ubiquitin (or empty plasmid) together with HA-NFATc3 in the presence or the absence of Flag-Trim39 or its SIM3 mutant, for 24 h. Then, cells were treated as in A. Ubiquitinated proteins and input lysates were analyzed as in A. The intensity of the ubiquitinated forms of NFATc3 was quantified and normalized by the intensity of NFATc3 in the total lysate. Relative values are indicated in red.

Article Snippet: Rabbit polyclonal antibody against Trim39 was from Proteintech (#12757-1-AP).

Techniques: Transfection, Ubiquitin Proteomics, Incubation, Purification, Western Blot, SDS Page, Plasmid Preparation, Immunoprecipitation, Recombinant, Comparison, Mutagenesis

Journal: bioRxiv

Article Title: Trim39 regulates neuronal apoptosis by acting as a SUMO-targeted E3 ubiquitin-ligase for the transcription factor NFATc3

doi: 10.1101/2020.09.29.317958

Figure Lengend Snippet: A. CGN primary cultures were transfected after 5 days in vitro (DIV 5) with GFP (as a negative control), WT GFP-NFATc3 or GFP-NFATc3-EallA for 16 h. Then, neurons were switched to serum-free medium containing 5 mM KCl (K5) for 7 h or were left untreated (control). Following fixation, nuclei were visualized by DAPI staining and proteins fused to GFP were detected by fluorescent microscopy. Arrows indicate GFP-positive neurons with thick arrows for neurons undergoing apoptosis and thin arrows for healthy neurons. B. The percentage of transfected, GFP-positive neurons undergoing apoptosis was assessed by examining cell morphology and nuclear condensation. Data are the means ± S.E.M. of four independent experiments performed as in A. * P <0.05; *** * P <0.001 significantly different from the corresponding value obtained in neurons transfected with GFP (two-way ANOVA followed by Sidak’s multiple comparisons test). C. CGNs were transduced with lentiviral particles expressing a non-targeting control (directed against Luciferase) or an shRNA specifically targeting Trim39 one day after plating. At DIV 6, total cell extracts from KCl-deprived neurons were analyzed by western blot using anti-Trim39 antibody (Origene). D. CGN were transduced and treated as in C. At DIV 6 they were incubated for 8 h in K5 medium, fixed and stained with DAPI. E. The percentage of apoptotic neurons was estimated by examining nuclear condensation. Data are the means ± S.E.M. of four independent experiments performed as in D. **** P <0.0001 significantly different from neurons transduced with the control shRNA (two-way ANOVA followed by Sidak’s multiple comparisons test).

Article Snippet: Rabbit polyclonal antibody against Trim39 was from Proteintech (#12757-1-AP).

Techniques: Transfection, In Vitro, Negative Control, Control, Staining, Microscopy, Transduction, Expressing, Luciferase, shRNA, Western Blot, Incubation

Journal: Arthritis Research & Therapy

Article Title: Potential therapeutic antibodies targeting specific adiponectin isoforms in rheumatoid arthritis

doi: 10.1186/s13075-018-1736-3

Figure Lengend Snippet: Detection of serum adiponectin isoforms with monoclonal antibodies (mAbs) against recombinant human adiponectin. a Western blot to screen mAbs detecting different adiponectin isoforms. Human serum was loaded into each lane of the gel and resolved by polyacrylamide gel electrophoresis (PAGE) and then transferred to a membrane. Each lane of the membrane was then separately cut and incubated with each mAb from hybridoma culture supernatant. After the secondary antibody was probed, the cut membrane was combined to form one sheet and developed with ECL solution. Eleven different mAbs from culture supernatant were tested to investigate their recognition patterns of adiponectin isoforms in serum. The mAb KH7–33, in lane 3, was selected as a representative recognizing only the middle molecular weight (MMW) isoform of adiponectin. The mAb KH7–41, in lane 4, was selected as a representative recognizing both the MMW and low molecular weight (LMW) isoforms of adiponectin. The mAb KH4–8, in lane 8, was selected as a representative mAb recognizing both the MMW and HMW isoforms of adiponectin. b Comparison of the mAb recognition patterns of adiponectin isoforms in human, mouse, and rat sera by Western blot. Human, mouse, and rat sera were separated by sodium dodecyl sulfate–PAGE. All three mAbs (KH7–33, KH7–41, and KH4–8) recognized the rat and mouse adiponectin MMW isoform. Lane 1, KH7–41; lane 2, KH7–33; lane 3, KH4–8; lane 4, commercial antibody against human adiponectin (Boster Immunoleader cat. no. PB9001) or mouse/rat adiponectin (Boster Immunoleader cat. no. PB9011). Mouse and rat serum were obtained via heart puncture of male BALB/c mice and Sprague Dawley rats (8 weeks old), respectively. Human serum was obtained from a male volunteer (55 years old)

Article Snippet: Lane 1, KH7–41; lane 2, KH7–33; lane 3, KH4–8; lane 4, commercial antibody against human adiponectin (Boster Immunoleader cat. no. PB9001) or mouse/rat adiponectin (Boster Immunoleader cat. no. PB9011).

Techniques: Bioprocessing, Recombinant, Western Blot, Polyacrylamide Gel Electrophoresis, Membrane, Incubation, Molecular Weight, Comparison

Journal: Arthritis Research & Therapy

Article Title: Potential therapeutic antibodies targeting specific adiponectin isoforms in rheumatoid arthritis

doi: 10.1186/s13075-018-1736-3

Figure Lengend Snippet: Immunohistological assays in human tissues with monoclonal antibodies (mAbs). To determine the pattern of recognition of adiponectin isoforms in human tissues by mAbs, a normal human adipose tissue was immunostained with mAbs (KH7–41, KH7–33, and KH4–8) (200×, scale bar = 25 μm). mAbs recognized adiponectin in the nucleus of adipocytes (blue arrow) and in vessels and endothelial cells (red arrow). b Human lung, kidney, and pancreas were stained with the KH7–41, KH7–33, and KH4–8 mAbs (100×). Abbreviations: HMW high molecular weight, LMW low molecular weight, MMW middle molecular weight

Article Snippet: Lane 1, KH7–41; lane 2, KH7–33; lane 3, KH4–8; lane 4, commercial antibody against human adiponectin (Boster Immunoleader cat. no. PB9001) or mouse/rat adiponectin (Boster Immunoleader cat. no. PB9011).

Techniques: Bioprocessing, Staining, High Molecular Weight, Molecular Weight

Journal: Arthritis Research & Therapy

Article Title: Potential therapeutic antibodies targeting specific adiponectin isoforms in rheumatoid arthritis

doi: 10.1186/s13075-018-1736-3

Figure Lengend Snippet: Inhibition of adiponectin-mediated gene expression in vitro by monoclonal antibodies (mAbs). To test the ability of the mAb KH4–8 to block adiponectin function, ( a ) human osteoblasts and ( b ) human umbilical vein endothelial cells (HUVECs) were treated with adiponectin (ADIPO) or KH4–8 mAb (mAb) or both. The mAb (~120 μg/mL) and recombinant adiponectin (2.5 μg/mL) were mixed and incubated for 1 h before being used to treat cells. After 24-h treatment, the culture supernatants were collected and frozen, and interleukin-6 (IL-6) and IL-8 were measured by using enzyme-linked immunosorbent assay (ELISA) (R&D Systems, Minneapolis, MN, USA). The experiments were performed in quadruplicate. The data shown are representative of three independent experiments, and similar results were obtained with all three mAbs. Values are expressed as mean ± standard error of the mean. The expression levels of the factors were compared between groups by using the Mann–Whitney test. * P <0.05, ** P <0.01 versus the untreated group, # P <0.05, ## P <0.01 versus the group treated with adiponectin and mAb

Article Snippet: Lane 1, KH7–41; lane 2, KH7–33; lane 3, KH4–8; lane 4, commercial antibody against human adiponectin (Boster Immunoleader cat. no. PB9001) or mouse/rat adiponectin (Boster Immunoleader cat. no. PB9011).

Techniques: Inhibition, Gene Expression, In Vitro, Bioprocessing, Blocking Assay, Recombinant, Incubation, Enzyme-linked Immunosorbent Assay, Expressing, MANN-WHITNEY

Journal: Arthritis Research & Therapy

Article Title: Potential therapeutic antibodies targeting specific adiponectin isoforms in rheumatoid arthritis

doi: 10.1186/s13075-018-1736-3

Figure Lengend Snippet: Epitope mapping of monoclonal antibody (mAb) KH4–8 against adiponectin. To identify the epitope-recognizing site of the KH4–8 mAb, PEPperMAP ® technology was performed as described in the Methods. Human adiponectin was translated into linear 15–amino acid peptides with a peptide-peptide overlap of 14 amino acids. Human adiponectin peptide microarrays were incubated with mouse mAb KH4–8 at different concentrations followed by staining with secondary goat anti-mouse IgG (H + L) DyLight680 antibody. The light intensity was read by a reader. The amino acid sequence QQNHYD (139–144) was confirmed from among the full 244–amino acid sequence to be the epitope of adiponectin

Article Snippet: Lane 1, KH7–41; lane 2, KH7–33; lane 3, KH4–8; lane 4, commercial antibody against human adiponectin (Boster Immunoleader cat. no. PB9001) or mouse/rat adiponectin (Boster Immunoleader cat. no. PB9011).

Techniques: Incubation, Staining, Sequencing

Journal: Arthritis Research & Therapy

Article Title: Potential therapeutic antibodies targeting specific adiponectin isoforms in rheumatoid arthritis

doi: 10.1186/s13075-018-1736-3

Figure Lengend Snippet: Anti-inflammatory effect of monoclonal antibodies (mAbs) KH4–8 and KH7–33 on the expression of serum pro-inflammatory cytokines of the collagen-induced arthritis mouse model. The serum levels of adiponectin, interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), and receptor activator of nuclear factor-kappa Β ligand (RANKL) were analyzed by using the Luminex system. Values are expressed as mean ± standard error of the mean. The expression levels of the factors were compared between groups ( n = 8) by using the Mann–Whitney test. ** P <0.01, * P <0.05 versus normal (NOR) group, and ## P <0.01, ## P <0.05 versus the control (CON) group. Abbreviations: ns not significant, pre prednisolone

Article Snippet: Lane 1, KH7–41; lane 2, KH7–33; lane 3, KH4–8; lane 4, commercial antibody against human adiponectin (Boster Immunoleader cat. no. PB9001) or mouse/rat adiponectin (Boster Immunoleader cat. no. PB9011).

Techniques: Bioprocessing, Expressing, Luminex, MANN-WHITNEY, Control

Journal: Arthritis Research & Therapy

Article Title: Potential therapeutic antibodies targeting specific adiponectin isoforms in rheumatoid arthritis

doi: 10.1186/s13075-018-1736-3

Figure Lengend Snippet: Anti-adiponectin antibodies reduce the histological signs of inflammation. The upper and lower panels present hematoxylin and eosin (H&E) staining and immunostaining against mouse adiponectin of mouse knee joints (n = 8), respectively. a Normal, b control, saline-treated arthritic, c KH7–33-treated arthritic, d KH4–8-treated arthritic, and ( e ) prednisolone-treated arthritic mice. Tissue structure was visualized by using H&E staining (original magnification, 40×). Scale bar = 2 mm. f Arthritic symptoms were evaluated by scoring the degree of inflammation on H&E histological sections of knee joints as described in the Methods. Small blue squares on H&E staining are magnified in the upper right corner (400×). Abbreviations: C cartilage, F femur, M meniscus, S subchondral bone, T tibia. In the lower panel, immunohistochemistry (IHC) reveals adiponectin expression in collagen-induced arthritis mouse joints (200×). The increased adiponectin expression level observed on IHC was not decreased by monoclonal antibody treatment. Adiponectin immunostaining score level was evaluated as described in the Methods. Results are presented as the mean of experiments (± standard error of the mean indicated by error bar) (one-way analysis of variance followed by Dunn’s multiple comparison test). *** P <0.001 versus the normal (NOR) group and # P <0.05, ## P <0.01 versus the control (CON) group. Abbreviation: pre prednisolone

Article Snippet: Lane 1, KH7–41; lane 2, KH7–33; lane 3, KH4–8; lane 4, commercial antibody against human adiponectin (Boster Immunoleader cat. no. PB9001) or mouse/rat adiponectin (Boster Immunoleader cat. no. PB9011).

Techniques: Staining, Immunostaining, Control, Saline, Immunohistochemistry, Expressing, Comparison

Journal: International Journal of Molecular Sciences

Article Title: Prolonged Chronic Consumption of a High Fat with Sucrose Diet Alters the Morphology of the Small Intestine

doi: 10.3390/ijms22147280

Figure Lengend Snippet: ( A , C ) Immunohistochemistry of adiponectin and adiponectin receptor of proximal tract of the small intestine. (Original magnification: 10×; scale bar: 100 μm). ( B , D ) The expression of adiponectin and its receptor was significantly reduced in HFD w/Suc mice compared to the SD group. * p < 0.05. These images are representative of n = 5 SD, n = 10 HFD w/Suc mice.

Article Snippet: Adiponectin , Boster Biological Technology, Pleasanton, CA, USA; code PA2014-1 , 1:50.

Techniques: Immunohistochemistry, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Prolonged Chronic Consumption of a High Fat with Sucrose Diet Alters the Morphology of the Small Intestine

doi: 10.3390/ijms22147280

Figure Lengend Snippet: Antibodies used with their sources and dilutions.

Article Snippet: Adiponectin , Boster Biological Technology, Pleasanton, CA, USA; code PA2014-1 , 1:50.

Techniques:

Journal: bioRxiv

Article Title: Ancestry Influences on the Molecular Presentation of Tumours

doi: 10.1101/2020.08.02.233528

Figure Lengend Snippet: (A) Summary of all detected ancestry-associated CNAs with numbers of gains (above x-axis) and losses (below x-axis) identified in each tumour context. Only tumour-types with at least one significant event are shown. (B) Pan-cancer ancestry-associations in CNAs for TCGA data. Each plot shows the logistic regression coefficient estimate (for age analyses) or difference in proportion (for ancestry analyses) for the indicated variable and CNA type. Dot colour indicates statistical significance, where red (copy number gain) and blue (copy number loss) show adjusted p < 0.05 and yellow (gain) and green (loss) show whether the multiple-testing adjusted p < 0.1 threshold is met. (C) Summary of ancestry-associated pan-cancer CNA drivers. Both TCGA and PCAWG findings are shown, and dot size indicates the effect-size as a proportion difference. Background shading shows multiple-testing adjusted multivariate p-values. The covariate to the right shows copy number gain drivers in red and loss drivers in blue. (D) EAN- and AFR-associations in TCGA kidney clear cell cancer CNAs are associated with (E) changes in mRNA abundance. The adjusted p-value is plotted against the coefficient of the CNA-age interaction for mRNA abundance, with each point representing a gene. Black dots show significant associations between mRNA and CNA; red dots show significant CNA-ancestry interactions. (F) RASSF1 and (G) MAP4 mRNA abundance changes between copy number loss (red) or no loss (black) in tumours of EUR and AFR ancestry. Adjusted CNA-AFR interaction p-value is shown. Tukey boxplots are depicted, as described in .

Article Snippet: For mRNA abundance, Illumina HiSeq rnaseqv2 level 3 RSEM normalised profiles were used.

Techniques:

Journal: bioRxiv

Article Title: Ancestry Influences on the Molecular Presentation of Tumours

doi: 10.1101/2020.08.02.233528

Figure Lengend Snippet: (A) Summary of all detected ancestry-associated SNVs found in each cancer type. Only tumour-types with at least one significant event shown. (B) pan-PCAWG and PCAWG tumour-type-specific ancestry-associations in driver and mitochondrial SNV frequency with the top showing adjusted multivariate p-values, middle showing difference in proportion, and bottom showing proportion of tumours with mutated gene per ancestry group. Covariate bars indicate tumour and element type context of each mutation, where coding sequence is abbreviated to CDS and mitochondrial DNA is mtDNA. (C) The 20 SNVs most associated with AFR and EAN ancestry in the TCGA colorectal and renal cancer (COADREAD) dataset. (D) Differential mRNA abundance associated with EAN-associated SNVs in TCGA COADREAD. Each point represents a gene with black dots showing significant associations between mRNA and SNV. (F) AXIN1 and (G) STK36 mRNA abundance changes between copy number loss (red) or no loss (black) compared by ancestry. Adjusted SNV term p-value is shown. Tukey boxplots are depicted, as described in .

Article Snippet: For mRNA abundance, Illumina HiSeq rnaseqv2 level 3 RSEM normalised profiles were used.

Techniques: Mutagenesis, Sequencing