Journal: Cancer cell

Article Title: MUTANT EZH2 INDUCES A PRE-MALIGNANT LYMPHOMA NICHE BY REPROGRAMMING THE IMMUNE RESPONSE

doi: 10.1016/j.ccell.2020.04.004

Figure Lengend Snippet: A. Mixed chimera mice (n=5) immunized with NP-OVA were treated with two doses of 100 µg lymphotoxin mLTβR-mIgG1 or control IgG antibody.

Article Snippet: In the experiments where interactions with Tfh or FDC were blocked in vivo , mice received 100 μg anti CD40L antibody i.v. (clone MR-1, BioXCell BE0017), 150 μg anti ICAM-1 antibody i.p. (clone YN1/1.7.4, BioXCell BE0020), 100 μg recombinant mLTβR (a fusion protein of lymphotoxin β receptor and Fc region of mouse IgG, which acts as inhibitor of transmembrane LTβR) i.v. (R&D Systems 1008-LR), or control IgG antibodies (BioXCell BE0091 and BE0090).

Techniques: Control

Journal: Cancer cell

Article Title: MUTANT EZH2 INDUCES A PRE-MALIGNANT LYMPHOMA NICHE BY REPROGRAMMING THE IMMUNE RESPONSE

doi: 10.1016/j.ccell.2020.04.004

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: In the experiments where interactions with Tfh or FDC were blocked in vivo , mice received 100 μg anti CD40L antibody i.v. (clone MR-1, BioXCell BE0017), 150 μg anti ICAM-1 antibody i.p. (clone YN1/1.7.4, BioXCell BE0020), 100 μg recombinant mLTβR (a fusion protein of lymphotoxin β receptor and Fc region of mouse IgG, which acts as inhibitor of transmembrane LTβR) i.v. (R&D Systems 1008-LR), or control IgG antibodies (BioXCell BE0091 and BE0090).

Techniques: Control, Blocking Assay, Recombinant, Adjuvant, Plasmid Preparation, Binding Assay, Staining, RNA Library Preparation, MicroChIP Assay, Sequencing, Microarray, Knock-In, Software, Gene Expression, Targeted Proteomics

Journal: BioMed Research International

Article Title: LRG1 May Accelerate the Progression of ccRCC via the TGF- β Pathway

doi: 10.1155/2020/1285068

Figure Lengend Snippet: The differential expression of LRG1 in ccRCC patients. (a) LRG1 is upregulated in primary tumor ( p < 0.001). (b) LRG1 expression of male patients is significantly different from male and normal contols ( p < 0.001). (c) LRG1 expression was significantly different between the normal control group and the 41-60 ( p = 0.0024) and 61-80-year-old subgroups ( p < 0.0001). (d) Caucasian group is the most differential expressed group compared with normal controls ( p < 0.0001). (e) LRG1 expression levels were significantly different between the normal control group and different ccRCC stage subgroups ( p < 0.0001). (f) All grades except grade 1 were significantly different from that in normal controls ( p < 0.05). ∗ ccRCC: clear cell renal cell carcinoma; TPM: transcript per million.

Article Snippet: The cytokines used in the study were carrier-free recombinant human LRG1 (R&D, 7890-LR).

Techniques: Quantitative Proteomics, Expressing, Control

Journal: BioMed Research International

Article Title: LRG1 May Accelerate the Progression of ccRCC via the TGF- β Pathway

doi: 10.1155/2020/1285068

Figure Lengend Snippet: Survival curves and differential methylation and expression of LRG1 promoter in ccRCC patients. (a) Low LRG1 expression indicates a prolonged patient survival time. The FPKM cutoff value of high expression and low expression is 1.19. (b) The methylation level of LRG1 gene has a strong negative correlation (corr = 0.677) with the expression of LRG1.

Article Snippet: The cytokines used in the study were carrier-free recombinant human LRG1 (R&D, 7890-LR).

Techniques: Methylation, Expressing

Journal: BioMed Research International

Article Title: LRG1 May Accelerate the Progression of ccRCC via the TGF- β Pathway

doi: 10.1155/2020/1285068

Figure Lengend Snippet: Promoter methylation level of LRG1 gene in ccRCC and subgroups. (a) Promoter methylation level of LRG1 gene is significantly downregulated compared with normal controls. (b) Methylation level of LRG1 gene in male and female patients is decreased compared with normal patients ( p < 0.0001). (c) Methylation level of LRG1 gene in different ages has significant differences compared with normal controls ( p < 0.0001). (d) Methylation level of LRG1 gene in different races has significant differences compared with normal controls ( p < 0.001). (e) Methylation level of all of the stages are downregulated than normal controls ( p < 0.001). (f) Methylation level of all of the grads are downregulated than normal controls ( p < 0.0001). (g) Methylation level of metastatic ccRCC is lower than nonmetastatic, but both of them are significantly downregulated than normal controls ( p < 0.001).

Article Snippet: The cytokines used in the study were carrier-free recombinant human LRG1 (R&D, 7890-LR).

Techniques: Methylation

Journal: BioMed Research International

Article Title: LRG1 May Accelerate the Progression of ccRCC via the TGF- β Pathway

doi: 10.1155/2020/1285068

Figure Lengend Snippet: Expression and methylation level in carcinoma and paracarcinoma samples of ccRCC. (a) mRNA expression of LRG1 in carcinoma and paracarcinoma tissues ( p < 0.01, n = 6) detected by qPCR. (b, c) Protein expression of LRG1 in carcinoma and paracarcinoma tissues ( p < 0.01, n = 6) detected by western blot. (d) LRG1 methylation level of CpG1 and CpG2 is downregulated in ccRCC tissue ( p < 0.0001).

Article Snippet: The cytokines used in the study were carrier-free recombinant human LRG1 (R&D, 7890-LR).

Techniques: Expressing, Methylation, Western Blot

Journal: BioMed Research International

Article Title: LRG1 May Accelerate the Progression of ccRCC via the TGF- β Pathway

doi: 10.1155/2020/1285068

Figure Lengend Snippet: Expression of TGFB1 in ccRCC patients. (a) Expression of TGFB1 in ccRCC patients is upregulated ( p < 0.001). (b) TGFB1 is upregulated in 789-O cells after stimulated with LRG1 ( p < 0.001) but not with heat-denatured LRG1. (c) TGFB1 is downregulated in 789-O cells after LRG1 knockdown ( p < 0.001).

Article Snippet: The cytokines used in the study were carrier-free recombinant human LRG1 (R&D, 7890-LR).

Techniques: Expressing, Knockdown

Journal: Journal of Biological Chemistry

Article Title: Bone Overgrowth-associated Mutations in the LRP4 Gene Impair Sclerostin Facilitator Function

doi: 10.1074/jbc.m110.190330

Figure Lengend Snippet: FIGURE 3. LRP4 is expressed in human bone, and Lrp4 silencing blunts sclerostin-mediated inhibition of in vitro bone mineralization. A, LRP4 protein is expressed in human osteoblasts (arrowheads) and osteocytes (arrows). Immunohistochemistry of LRP4 in human femoral neck from a male subject aged 75 is shown. Scale bar, 50 m. B, LRP4 RNA is expressed in samples from human femoral neck. RNA was extracted from a female subject aged 65 and a male subject aged 80. PTH1R, LRP4, SOST, LRP5, and LRP6 RNA levels were assessed by real-time quantitative PCR and normalized with GAPDH, and relative expression compared with PTH1R is shown. C, knockdown of LRP4 in Kusa-A1 blocks the inhibitory effect of sclerostin on in vitro mineralization activity. Kusa-A1 cells were transducedwithlentiviralparticlesharboringshRNAagainstLrp4.ReductionofLrp4RNAlevelswasassessedbyreal-timequantitativePCR,priortotheaddition of an osteoblastic differentiation medium. Calcium content was assessed 4 days later. Error bars, S.E.

Article Snippet: Recombinant human DKK1 and LRP6 were purchased from R&D Systems.

Techniques: Inhibition, In Vitro, Immunohistochemistry, Real-time Polymerase Chain Reaction, Expressing, Knockdown, Activity Assay

Journal: EBioMedicine

Article Title: Molecular basis of reduced LAIR1 expression in childhood severe malarial anaemia: Implications for leukocyte inhibitory signalling

doi: 10.1016/j.ebiom.2019.06.040

Figure Lengend Snippet: Relationship between LAIR1 expression and intraleukocytic haemozoin. (a) Pairwise comparisons of LAIR1 transcript levels between children with and without pigment-containing monocytes (PCM) and pigment-containing neutrophils (PCN) in the non-SMA and SMA groups were determined using Mann-Whitney U test. Data are presented as box-plots, where the box represents the interquartile range, the line through the box is the median, and whiskers show the 10th and 90th percentiles. LAIR1 transcript levels were lower in children with PCM compared to those without, while LAIR1 transcript levels were comparable between absence/presence of PCN. (b) Temporal kinetics of LAIR1 transcript levels in response to Pf Hz treatment of PBMC from malaria-naïve donors ( n = 6, measured in triplicate). Pairwise comparisons were determined using Student t -test. Significant ( P < .05) differences in transcript levels between no treatment and Pf Hz treatment (10 μg/mL) groups represented by *at 0.5, 2, and 4 h time points. Data represent average of individuals ( n = 3) with each condition performed in triplicate (error bars represent SEM). (c) Immunoblot analysis of non-treated (baseline) and Pf Hz-treated (10 μg/mL) PBMC lysates for 12, 24, and 48 h. (d) Densitometric analysis of normalised cellular LAIR1 protein production presented as mean ± SEM. Across group comparisons analysed using ANOVA. Pairwise comparisons analysed using Student t-test. Data represent average of individuals ( n = 6) with each condition performed in triplicate (error bars represent SEM). LAIR1 protein levels were lower in Pf Hz-treated PBMC lysates compared to no treatment at 12, 24, and 24 h.

Article Snippet: Soluble LAIR1 was measured in serum of study participants using the human LAIR1 ELISA matched antibody pair, and recombinant human LAIR1 (Creative Diagnostics, 45–16 Ramsey Rd, Shirley, NY 11967; Cat No. ABPR-0519).

Techniques: Expressing, MANN-WHITNEY, Western Blot

Journal: EBioMedicine

Article Title: Molecular basis of reduced LAIR1 expression in childhood severe malarial anaemia: Implications for leukocyte inhibitory signalling

doi: 10.1016/j.ebiom.2019.06.040

Figure Lengend Snippet: Effect of intraleukocytic Pf Hz on LAIR1 and SHP-1 phosphorylation. Temporal kinetics of the signalling pathway in response to the different treatment conditions was determined in PBMCs from malaria-naïve donors (n = 6, measured in triplicate). (a) Human Phospho-immunoreceptor array results showing LAIR1 and SHP-1 phosphorylation upon no treatment (baseline), C1q (25 μg/mL), Pf Hz (10 μg/mL), and Pf Hz + C1q. Phosphorylation (spots) in the different conditions with control represented by blue boxes, LAIR1 by red boxes, and SHP-1 by green boxes. (b) Densitometric analysis of human phosphor-immunoreceptor array data. Data presented as (mean ± SEM). Across group comparisons analysed using ANOVA. *indicates significant differences ( P < .05) determined by Student t-test in pLAIR1 and pSHP-1 compared to C1q treatment. Phagocytosis of Pf Hz resulted in a reduction of pLAIR1 relative to C1q (alone) treatment. Similarly, ingestion of Pf Hz also caused a marked decrease in pSHP-1 levels relative to C1q treatment. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Soluble LAIR1 was measured in serum of study participants using the human LAIR1 ELISA matched antibody pair, and recombinant human LAIR1 (Creative Diagnostics, 45–16 Ramsey Rd, Shirley, NY 11967; Cat No. ABPR-0519).

Techniques:

Journal: EBioMedicine

Article Title: Molecular basis of reduced LAIR1 expression in childhood severe malarial anaemia: Implications for leukocyte inhibitory signalling

doi: 10.1016/j.ebiom.2019.06.040

Figure Lengend Snippet: LAIR1 signalling pathway in malaria. The LAIR1 pathway is activated by attachment of collagen and collagenous ligands (C1q) to LAIR1 extracellular surface. This results in phosphorylation of intracellular LAIR1 ITIM tyrosine residues by Src family kinases. Phosphorylated ITIMs serve as docking sites for recruitment of SHP-1 and SHP-2 phosphatases. SHP-1 and SHP-2 become localized to phosphorylated ITIMs through their regulatory SH2 domains, subsequently inducing their phosphatase activity. Activated SHP-1 has been shown to block activation and nuclear translocation of nuclear factor nuclear factor-kappa beta (NF-κB) through de-phosphorylation of inhibitor of kappa-beta kinase complex (IKK). SHP-1 phosphatase activity also inhibits activation and translocation of IRFs from the cytoplasm to the nucleus by preventing TANK-binding kinase 1 (TANK-1) phosphorylation of interferon regulatory factors (IRFs). These events subsequently block transcription of inflammatory mediator encoding genes with response elements for IRFs and NF-κB. SHP-2 inhibits activation of IRF 8 and blocks expression of phagocyte NADPH oxidase (gp91 PHOX ). LAIR1 inhibitory signalling is regulated by soluble LAIR1 and LAIR2 (soluble homolog of LAIR1) through competition for collagenous ligands. Children with SMA had increased circulating levels of sLAIR1 and sLAIR2 indicative of enhanced receptor shedding. C1q was reduced in children with SMA, thereby, limiting ligand availability. Phagocytosis of Pf Hz antagonizes LAIR1 signalling through down-regulation of LAIR1 ITIM and SHP-1 phosphorylation, but does not alter SHP-2 phosphorylation. Leukocytic ingestion of Pf Hz also decreases LAIR1 transcripts and protein. These events result in NF-κB activation and the consequent production of pro-inflammatory mediators that enhance the pathogenesis of SMA. Red arrows represent ligand-receptor interaction, black solid arrows represent activation, and dashed black arrows represent blockade. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Soluble LAIR1 was measured in serum of study participants using the human LAIR1 ELISA matched antibody pair, and recombinant human LAIR1 (Creative Diagnostics, 45–16 Ramsey Rd, Shirley, NY 11967; Cat No. ABPR-0519).

Techniques: Activity Assay, Blocking Assay, Activation Assay, Translocation Assay, De-Phosphorylation Assay, Binding Assay, Expressing

Journal: Cell

Article Title: Mitophagy in Intestinal Epithelial Cells Triggers Adaptive Immunity during Tumorigenesis

doi: 10.1016/j.cell.2018.05.028

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Assay was performed in triplicates in a reaction buffer (50 mM sodium acetate, 8 mM EDTA, 1 mM DTT, 1 μM pefabloc, all Sigma-Aldrich) using 2 μg of protein lysate and 10 μM protease substrate z-LR-AMC (ES008, R&D Systems).

Techniques: In Vivo, Flow Cytometry, Recombinant, Staining, Cell Isolation, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Glucose Assay, Reverse Transcription, SYBR Green Assay, Control, Software

Journal: Journal for immunotherapy of cancer

Article Title: Selective targeting of GARP-LTGFβ axis in the tumor microenvironment augments PD-1 blockade via enhancing CD8 + T cell antitumor immunity.

doi: 10.1136/jitc-2022-005433

Figure Lengend Snippet: Figure 2 In vitro characterization of anti-GARP antibody PIIO-1. (A) GARP expression on human regulatory T cells and platelets was evaluated by flow cytometry after staining with PIIO-1 at 10 µg/mL. (B) 293 FT cell line was transfected with empty vector (EV), human GARP (hGARP)-expressing vector only or co-transfected with hGARP and latent TGFβ1 expression vectors. GARP expression on indicated cell line was detected by flow cytometry after staining with PIIO-1 at 10 µg/mL. (C) Human GARP sequence was replaced by murine GARP according to the schematic diagram to generate the chimeric constructs of human and murine GARP that were tagged with HA (hemagglutinin) epitope. Transfection efficiency was determined using anti- HA antibody. All constructs were transfected into 293 FT cells. (D) Crystal structure of the GARP (green)-LTGFβ (gray) complex (PDB DOI: 10.2210/pdb6GFF/pdb). The region of PIIO-1 recognition is orange and the residues interacting with LTGFβ are cyan. LTGFβ occludes approximately 30% of the potential antibody binding site and may sterically or allosterically restrict access of the antibody to GARP in the LTGFβ-complexed state. Modeling was carried out using Pymol. (E) Jurkat cell line, made to overexpress hGARP, was incubated with LTGFβ1 along with mIgG1 or PIIO-1 at indicated concentration for 30 min at 37℃. Human LAP expression was detected by flow cytometry. All data are representative of 2–6 independent experiments. GARP, Glycoprotein-A repetitions predominant; LAP, latency-associated peptide.

Article Snippet: Generation of anti-human GARP (hGARP) antibodies The generation of anti- hGARP antibody has been described previously.19 BALB/c mice was immunized with recombinant human GARP (R&D Systems) in Freund’s complete adjuvant and followed by boosting with SP2/0hGARP cells for 2–3 times.

Techniques: In Vitro, Expressing, Flow Cytometry, Staining, Transfection, Plasmid Preparation, Sequencing, Construct, Binding Assay, Incubation, Concentration Assay

Journal: Journal for immunotherapy of cancer

Article Title: Selective targeting of GARP-LTGFβ axis in the tumor microenvironment augments PD-1 blockade via enhancing CD8 + T cell antitumor immunity.

doi: 10.1136/jitc-2022-005433

Figure Lengend Snippet: Figure 3 PIIO-1 enhanced antitumor efficacy of anti-PD-1 ICB in GARP+ TNBC. (A) Experimental scheme. BALB/c mice were injected with 1×105 4T1-hGARP cells in the mammary fat pad, followed by i.p. injection of 200 µg/mouse of PIIO-1 and/or 150 µg/mouse anti-PD-1 every 3 days. (B) Primary tumor growth curve. (C) Overall survival of mice. (D) Summary of the incidence of tumor-free mice among groups. (E) Lungs were collected and sectioned at experimental end point, then stained with H&E. Representative images from each group of mice are shown. Scale bar, 20 µm. The numbers of visible lung metastatic nodules are quantified. (F) Summary of the incidence of metastasis among groups. (G) Tumors were collected at end point and stained by IHC for pSMAD3, α-SMA. Representative images of tumor tissues from four groups of mice are shown (left). Scale bar, 50 µm. Quantification of the IHC staining is shown (right). (H) Sera from each mouse was collected at end point. Total and active TGFβ level in the sera were assessed by ELISA. (I) Mice with tumor regression following combination therapy were monitored for 300 days, then rechallenged with 5×105 wild-type 4T1 mammary tumor in contralateral mammary fat pad. Naive BALB/c mice without pre-exposure to tumor were used as control. Shown is the overall survival. Tumor curve analysis was performed using repeated measures 2-way analysis of variance. Overall survival is analyzed by log-rank (Mantel-Cox) test. (E, G) were analyzed by paired t-test according to the tumor collection time points. Other data were analyzed by two-tailed Student’s t-test. B, C were corrected for multiple testing using the Tukey procedure. All data are presented as mean±SEM. *p<0.05, **p<0.005, ***p<0.001, ****p<0.0001.

Article Snippet: Generation of anti-human GARP (hGARP) antibodies The generation of anti- hGARP antibody has been described previously.19 BALB/c mice was immunized with recombinant human GARP (R&D Systems) in Freund’s complete adjuvant and followed by boosting with SP2/0hGARP cells for 2–3 times.

Techniques: Injection, Staining, Immunohistochemistry, Enzyme-linked Immunosorbent Assay, Control, Two Tailed Test

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Intranasal Delivery of N-terminal Modified Leptin-Pluronic Conjugate for Treatment of Obesity

doi: 10.1016/j.jconrel.2017.03.029

Figure Lengend Snippet: (a) MALDI-TOF spectra of reaction mixture. The ratio denotes the molar ratio of leptin monomer to polymer. (b) SEC of leptin conjugates eluted from Superdex75 100/300 GL column in 10% MeOH/0.25 M sodium phosphate, pH 7.5 at 0.5 ml/min. (c) SDS-PAGE of pooled fractions in SEC denoted by elution time. Fraction of 20.3–21.9 min in LepNP85 and fraction of 18.5–21 min in LepNPEG5K were used.

Article Snippet: Recombinant mouse leptin and recombinant human leptin receptor-Fc chimeras were purchased from R&D Systems (Minneapolis, MN).

Techniques: Polymer, SDS Page

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Intranasal Delivery of N-terminal Modified Leptin-Pluronic Conjugate for Treatment of Obesity

doi: 10.1016/j.jconrel.2017.03.029

Figure Lengend Snippet: SDS-PAGE of the reaction mixture of leptin and Leptin-2PCA with CHO-P85-OH or with P85-DSP.

Article Snippet: Recombinant mouse leptin and recombinant human leptin receptor-Fc chimeras were purchased from R&D Systems (Minneapolis, MN).

Techniques: SDS Page

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Intranasal Delivery of N-terminal Modified Leptin-Pluronic Conjugate for Treatment of Obesity

doi: 10.1016/j.jconrel.2017.03.029

Figure Lengend Snippet: (a) Leptin 1:1 physical mixture with P85 or PEG5K. (b) Leptin 1:1 conjugates.

Article Snippet: Recombinant mouse leptin and recombinant human leptin receptor-Fc chimeras were purchased from R&D Systems (Minneapolis, MN).

Techniques:

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Intranasal Delivery of N-terminal Modified Leptin-Pluronic Conjugate for Treatment of Obesity

doi: 10.1016/j.jconrel.2017.03.029

Figure Lengend Snippet: Leptin and leptin 1:1 conjugates were eluted from Jupiter C4 column (particle diameter 5 μm, pore diameter 300 Å, 4.6 × 100 mm) by gradient elution at 1 ml/min and 25 °C, and monitored by absorption at 220 nm. Mobile phase A: water + 0.1% TFA; mobile phase B: acetonitrile + 0.1% TFA: isopropanol 50:50 (v/v). The elution started from 5% B for 5 min, then linearly increased to 95% B at 1%/min, and stayed at 95% B till 100 min.

Article Snippet: Recombinant mouse leptin and recombinant human leptin receptor-Fc chimeras were purchased from R&D Systems (Minneapolis, MN).

Techniques:

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Intranasal Delivery of N-terminal Modified Leptin-Pluronic Conjugate for Treatment of Obesity

doi: 10.1016/j.jconrel.2017.03.029

Figure Lengend Snippet: (a) Representative sensorgram for native leptin. (b) Representative sensorgram for LepNP85 1:1 conjugates. (c) Representative sensorgram for LepDSSP85 1:1 conjugates. (d) Dissociation constants (KD) of leptin and leptin 1:1 conjugates. Data are mean ± SD, n= 5~12. *** p < 0.001 and n.s. not significant by One-way ANOVA and post Newman-Keuls Multiple Comparison Test.

Article Snippet: Recombinant mouse leptin and recombinant human leptin receptor-Fc chimeras were purchased from R&D Systems (Minneapolis, MN).

Techniques: Comparison

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Intranasal Delivery of N-terminal Modified Leptin-Pluronic Conjugate for Treatment of Obesity

doi: 10.1016/j.jconrel.2017.03.029

Figure Lengend Snippet: (a) Dose response of leptin and LepNP85 1:1 conjugates. Data are mean ± SEM, n=3. (b) Comparison of leptin and leptin 1:1 conjugates at 100 ng/mouse. Data are mean ± SEM, n= 5~7. ** p < 0.01 and *** p < 0.001 by One-way ANOVA and post Newman-Keuls Multiple Comparison Test.

Article Snippet: Recombinant mouse leptin and recombinant human leptin receptor-Fc chimeras were purchased from R&D Systems (Minneapolis, MN).

Techniques: Comparison

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Intranasal Delivery of N-terminal Modified Leptin-Pluronic Conjugate for Treatment of Obesity

doi: 10.1016/j.jconrel.2017.03.029

Figure Lengend Snippet: Serum clearance, unidirectional brain influx rates and initial volumes of brain distribution for leptin and leptin 1:1 conjugates.

Article Snippet: Recombinant mouse leptin and recombinant human leptin receptor-Fc chimeras were purchased from R&D Systems (Minneapolis, MN).

Techniques:

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Intranasal Delivery of N-terminal Modified Leptin-Pluronic Conjugate for Treatment of Obesity

doi: 10.1016/j.jconrel.2017.03.029

Figure Lengend Snippet: Male CD-1 mice were co-injected with 125I-LepNP85 and 131I-leptin. Data are mean ± SEM, n=7/time point, * p < 0.05, ** p < 0.01, and *** p < 0.001 by Two-way ANOVA.

Article Snippet: Recombinant mouse leptin and recombinant human leptin receptor-Fc chimeras were purchased from R&D Systems (Minneapolis, MN).

Techniques: Injection

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Intranasal Delivery of N-terminal Modified Leptin-Pluronic Conjugate for Treatment of Obesity

doi: 10.1016/j.jconrel.2017.03.029

Figure Lengend Snippet: Each male CD-1 mouse was injected with 125I-labeled LepNP85 with or without 10 μg of cold leptin. Data are mean ± SEM, n=7/time point, * p < 0.05 by Two-way ANOVA.

Article Snippet: Recombinant mouse leptin and recombinant human leptin receptor-Fc chimeras were purchased from R&D Systems (Minneapolis, MN).

Techniques: Injection, Labeling

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Intranasal Delivery of N-terminal Modified Leptin-Pluronic Conjugate for Treatment of Obesity

doi: 10.1016/j.jconrel.2017.03.029

Figure Lengend Snippet: 1 μg of biotin-labeled leptin and LepNP85 in 1 μl of PBS were injected locally into brain by ICV injection. The concentrations of biotin-labeled proteins in brain lysate and plasma were analyzed by ELISA. (a) Blood absorption of biotin-leptin and biotin-LepNP85 after ICV injection. (b) Brain retention of biotin-leptin and biotin-LepNP85 after ICV injection. Data are mean ± SEM, n=4/time point, * p < 0.05 by One-way ANOVA and post Dunnett’s Multiple Comparison Test, and *** p < 0.001 by Two-way ANOVA.

Article Snippet: Recombinant mouse leptin and recombinant human leptin receptor-Fc chimeras were purchased from R&D Systems (Minneapolis, MN).

Techniques: Labeling, Injection, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Comparison