Journal: bioRxiv

Article Title: Vascular endothelial growth factor-D improves lung vascular integrity during acute lung injury

doi: 10.1101/2024.12.16.628787

Figure Lengend Snippet: A) NICHES analysis of AFs, mesothelial cells, pericytes, gCap, and aCap in human microvascular niche yields a quantitative cell-cell signaling atlas of AF-aCap, AF-gCap, Mesothelial-aCap, Mesothelial-gCap, Pericyte-aCap, and Pericyte-gCap visualized by low-dimensional UMAP embedding. B) FeaturePlot of top differentially expressed L/R pairs ANGPT1 - TEK , SLIT2 - ROBO4 , VEGFD - KDR , BMP5-BMPR2 , ADM - RAMP2 , WNT5A - FZD6 , PTN - RACK1 , OXT - EDNRB , and ADCYAP1 - VIPR1 from NICHES analysis C) Top 10 differentially expressed L/R pairs between different cell-cell interaction signals processed through NICHES and plotted in DoHeatmap. D) Immunostaining of representative L/R pairs from AF to microvascular endothelial cells, including SLIT2 - ROBO4 , ADM - RAMP2 , BMP5 - BMPR2 , and VEGFD - VEGFR2 in human lung section. ITGA8 is a marker for AF; PRX is a marker for microvascular endothelial cells. Scale bar 20 μm.

Article Snippet: The protein levels of SLIT2, ADM, BMP5, and VEGFD in samples harvested from different timepoints were determined using corresponding ELISA kits: SLIT2 (# NB030384), ADM (# NBP2-78738), BMP5 (# NBP2-69996), VEGFD (# NBP2-78890), all from Novus Biologicals.

Techniques: Immunostaining, Marker

Journal: bioRxiv

Article Title: Vascular endothelial growth factor-D improves lung vascular integrity during acute lung injury

doi: 10.1101/2024.12.16.628787

Figure Lengend Snippet: A) C57Bl/6 mice were injected with LPS for 18 hours, followed by a collection of bronchioalveolar lavage (BAL) fluid. The expression level of VEGF-D was measured in the BAL fluid in mice with or without LPS challenge. B) Mice were injected with LPS, followed by treatment with VEGF-D (225 μg/kg). The BAL fluid and histological samples were collected after 18 hours of treatment. C) The FITC-albumin probe was injected retro-orbitally 16 hours after the LPS challenge. The amount of probe in BAL fluid was quantified after another 2 hours. The FITC-albumin levels in BAL fluid in the control and experimental groups were determined. D) The BAL fluid was spun down at 5,000 g for 10 minutes and resuspended in 300 μL of 1XPBS. The number of cells in the BAL from different conditions was determined using a hemacytometer. n = 12 – 14 per group. E) LPS and LPS+VEGFD lungs were excised, fixed in 4% paraformaldehyde, embedded in paraffin, and used for histochemical analysis after haematoxylin and eosin staining. Images are representative of 6–9 lung specimens for each condition. Scale bar 150 μm. F) Lung pathological index was performed by a blinded reviewer from 1 to 4, where 4 was the worst pathology. The score was used for both peribronchial and alveolar inflammation. The average of two scores was taken to obtain a range between 1 (no inflammation or pathology) and 4 (maximum inflammation and pathology). The lung pathological index was calculated in LPS and LPS+VEGFD conditions. G-I) The expression level of TNF-α and IL-6 were measured in BAL fluid from LPS and LPS+VEGFD conditions using corresponding ELISA kits. Mice without LPS challenge were used as control. N = 12 - 14 per treatment group. Data presented as mean ± SEM. * and ** indicate p<0.05 and p<0.01, respectively.

Article Snippet: The protein levels of SLIT2, ADM, BMP5, and VEGFD in samples harvested from different timepoints were determined using corresponding ELISA kits: SLIT2 (# NB030384), ADM (# NBP2-78738), BMP5 (# NBP2-69996), VEGFD (# NBP2-78890), all from Novus Biologicals.

Techniques: Injection, Expressing, Control, Staining, Enzyme-linked Immunosorbent Assay

Journal: Cell reports

Article Title: Impaired islet function and normal exocrine enzyme secretion occur with low inter-regional variation in type 1 diabetes

doi: 10.1016/j.celrep.2024.114346

Figure Lengend Snippet:

Article Snippet: Human Pancreatic Lipase ELISA Kit , Novus Biologicals , Cat# NB016815.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Software, Microscopy

Journal: iScience

Article Title: Amitriptyline revitalizes ICB response via dually inhibiting Kyn/Indole and 5-HT pathways of tryptophan metabolism in ovarian cancer

doi: 10.1016/j.isci.2024.111488

Figure Lengend Snippet: Expression of MYD88, IDO1, and IL4-I1 in EOC cells after LPS-induced combination with AMI (A−C) RT-qPCR and Western blot verification of MYD88 and IDO1 expression in A2780, OVCAR3, and ID8 EOC cells after LPS-induction (1 μg/ml). (D−F) mRNA expression of MYD88 and IDO1 by the combination of LPS with AMI in A2780, OVCAR3 (AMI: 50, 100μm) and ID8 (AMI: 30, 60μm) EOC cells. (G−H) MYD88 and IDO1 expression was not detected in A2780 EOC cells according to the pre-WB results. Therefore, only the expression of MYD88 and ID8 protein levels in OVCAR3 and ID8 were performed in the combined treatment. (Protein expression of MyD88 and IDO1 relative to Tubulin in EOC cells treated with different concentrations of drugs). (I) mRNA expression of IL4I1 on A2780, OVCAR3, and ID8 EOC cells after LPS combined with AMI treatment was analyzed by RT-qPCR, in which IL4I1, as a secreted protein, was detected in the levels in the EOC cells culture supernatant with Elisa. Each assay was performed in triplicate, and data are presented as mean ± SD ( n = 3).

Article Snippet: ELISA kit- TNF-α (Mouse) , Novus Biologicals , VAL609.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: Amitriptyline revitalizes ICB response via dually inhibiting Kyn/Indole and 5-HT pathways of tryptophan metabolism in ovarian cancer

doi: 10.1016/j.isci.2024.111488

Figure Lengend Snippet: The killing activity of T lymphocytes against EOC cells and expression of CD39 after AMI stimulation (A) Expression of CD8 + CD39 + in T lymphocytes after exposure to different concentrations of AMI-treated (AMI concentrations refer to the preceding) EOC cells culture supernatants for 12 or 24 h as analyzed by FCM. (B) The killing activity of corresponding T lymphocytes (effector cells) to parental EOC cells (target cells) or AMI-treated EOC cells (target cells) shown are from the effector/target ratio at 1:1, 5:1, and 10:1. Supernatant 0h, 24h, and 48h represent the stimulation of T lymphocytes by supernatants collected at different times from EOC cells treated with AMI, respectively. (C) Measure the levels of TNF-α and IFN-γ in lymphocyte culture supernatants stimulated with AMI-treated EOC cell culture supernatants by Elisa assays. Each assay was performed in triplicate, and data are presented as mean ± SD ( n = 3). In Elisa results, ∗ indicates compared to the lymphocytes without any treatment (LN), # indicates compared to the lymphocytes stimulated with DMSO-treated EOC cell culture supernatant. #, ##, ### and #### indicates p < 0.05, p < 0.01, p < 0.001, and p < 0.0001 respectively.

Article Snippet: ELISA kit- TNF-α (Mouse) , Novus Biologicals , VAL609.

Techniques: Activity Assay, Expressing, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: Amitriptyline revitalizes ICB response via dually inhibiting Kyn/Indole and 5-HT pathways of tryptophan metabolism in ovarian cancer

doi: 10.1016/j.isci.2024.111488

Figure Lengend Snippet: AMI reshapes TIME, the serum level of IL-4I1 and TRP metabolites peripheral blood of EOC-bearing mice (A and B) Tryptophan metabolites in the serum of unimplanted tumor-bearing mice (normal mice) and compared with implanted tumor-bearing mice (controls). (C and D) Tryptophan metabolites in the serum of normal mice (without implanted tumors) before and after the use of AMI. (E−H) EOC-bearing mice receiving PD-1 inhibitor - combined AMI therapy, LC-MS/MS was used to detect the amount of KYN, KYNA, TRP, and the ratio of KYN/TRP in the serum of EOC-bearing mice. (I) IL4I1 in EOC-bearing mice serum by Elisa. (J) LC-MS/MS was used to detect the amount of AHR ligands (I3P, IAA, I3A, and ILA) in the serum of EOC-bearing mice. Data are presented as mean ± SD ( n = 3).

Article Snippet: ELISA kit- TNF-α (Mouse) , Novus Biologicals , VAL609.

Techniques: Liquid Chromatography with Mass Spectroscopy, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: Amitriptyline revitalizes ICB response via dually inhibiting Kyn/Indole and 5-HT pathways of tryptophan metabolism in ovarian cancer

doi: 10.1016/j.isci.2024.111488

Figure Lengend Snippet:

Article Snippet: ELISA kit- TNF-α (Mouse) , Novus Biologicals , VAL609.

Techniques: Recombinant, Bicinchoninic Acid Protein Assay, Membrane, Isolation, Enzyme-linked Immunosorbent Assay, Lactate Dehydrogenase Assay, Software

Journal: Frontiers in Physiology

Article Title: The effect of 60 days of 6° head-down-tilt bed rest on circulating adropin, irisin, retinol binding protein-4 (RBP4) and individual metabolic responses in young, healthy males

doi: 10.3389/fphys.2024.1435448

Figure Lengend Snippet: Favourable metabolic actions of adropin and irisin and deleterious metabolic actions of retinol binding protein-4 (RBP4), when concentrations of RBP4 are elevated. Abbreviations: ER, endoplasmic reticulum; HGP, hepatic glucose production; JNK, c-Jun N-terminal Kinase; TG, triglycerides; WAT, white adipose tissue.

Article Snippet: Specifically, adropin was quantified with the Novus Biologicals Human Adropin ELISA (Cat# NBP2-66433; intra-assay CV: 13.61%), irisin was quantified using the BioVendor Human Irisin ELISA (Cat# RAG018R; intra-assay CV: 8.57%) and RBP4 was quantified using the BioVendor Human RBP4 ELISA (Cat# RAG005R; intra-assay CV: 8.93%).

Techniques: Binding Assay

Journal: Frontiers in Physiology

Article Title: The effect of 60 days of 6° head-down-tilt bed rest on circulating adropin, irisin, retinol binding protein-4 (RBP4) and individual metabolic responses in young, healthy males

doi: 10.3389/fphys.2024.1435448

Figure Lengend Snippet: The effect of 60 days HDT bed rest on circulating adropin (A) , irisin (B) , and RBP4 (C) measured on BDC-5 (pre) and HDT59 (post). Data are presented as mean ± SD. The results for adropin, irisin, and RBP4 (figure) include n = 11 in CTRL and n = 12 JUMP. Biomarker concentrations were corrected for changes in hemoconcentration post-HDT bed rest. Abbreviations: CTRL, control group; JUMP, jumping countermeasure group. A significant main effect of time indicated as * p ≤ 0.05 or *** p ≤ 0.001.

Article Snippet: Specifically, adropin was quantified with the Novus Biologicals Human Adropin ELISA (Cat# NBP2-66433; intra-assay CV: 13.61%), irisin was quantified using the BioVendor Human Irisin ELISA (Cat# RAG018R; intra-assay CV: 8.57%) and RBP4 was quantified using the BioVendor Human RBP4 ELISA (Cat# RAG005R; intra-assay CV: 8.93%).

Techniques: Biomarker Discovery, Control

Journal: Frontiers in Physiology

Article Title: The effect of 60 days of 6° head-down-tilt bed rest on circulating adropin, irisin, retinol binding protein-4 (RBP4) and individual metabolic responses in young, healthy males

doi: 10.3389/fphys.2024.1435448

Figure Lengend Snippet: Changes in circulating adropin (A) , irisin (B) , and RBP4 (C) when subjects were divided into two subgroups based on a decrease or an increase in insulin sensitivity post-HDT bed rest. Data are presented as mean ± SD. Metabolic characteristics were measured on BDC-5 and HDT59. Pre-to post-HDT bed rest changes in adropin, irisin, and RBP4 are shown in figure, respectively. Biomarker concentrations were corrected for changes in hemoconcentration post HDT-bed rest. Abbreviations: RBP4, retinol binding protein 4. The results for these circulating biomarkers include n = 17 in the decreased insulin sensitivity subgroup and n = 6 in the increased insulin sensitivity subgroup. * p ≤ 0.05 or ** p ≤ 0.01.

Article Snippet: Specifically, adropin was quantified with the Novus Biologicals Human Adropin ELISA (Cat# NBP2-66433; intra-assay CV: 13.61%), irisin was quantified using the BioVendor Human Irisin ELISA (Cat# RAG018R; intra-assay CV: 8.57%) and RBP4 was quantified using the BioVendor Human RBP4 ELISA (Cat# RAG005R; intra-assay CV: 8.93%).

Techniques: Biomarker Discovery, Binding Assay

Journal: Cancers

Article Title: New Advances in the Study of CMTM6, a Focus on Its Novel Non-Canonical Cellular Locations, and Functions beyond Its Role as a PD-L1 Stabilizer

doi: 10.3390/cancers16183126

Figure Lengend Snippet: Description of the cohort subjects (CMTM6).

Article Snippet: Human PD-L1 ELISA Kit, Lot 356381-001, REF BMS2327 (ThermoFisher, Waltham, MA, USA) and HUMAN CMTM6 ELISA Kit (Colorimetric), CAT NBP2-75298 (NOVUS BIOLOGICALS, Centennial, CO, USA) were used.

Techniques: Histopathology

Journal: Cancers

Article Title: New Advances in the Study of CMTM6, a Focus on Its Novel Non-Canonical Cellular Locations, and Functions beyond Its Role as a PD-L1 Stabilizer

doi: 10.3390/cancers16183126

Figure Lengend Snippet: Total CMTM6 and total PD-L1 in plasma from the healthy donors (HDs) group and the cervical cancer patients (CC) group. ( a ) Concentrations of CMTM6 in the plasma of HDs ( n = 23) and CC ( n = 23) were measured by enzyme-linked immunosorbent assay (ELISA). Data are shown as pg/mL of CMTM6. ( b ) PD-L1 levels in the plasma of HDs ( n = 11) and CC ( n = 21) are shown as pg/mL of PD-L1. Student’s t -test was used. * p ≤ 0.05, ** p ≤ 0.01.

Article Snippet: Human PD-L1 ELISA Kit, Lot 356381-001, REF BMS2327 (ThermoFisher, Waltham, MA, USA) and HUMAN CMTM6 ELISA Kit (Colorimetric), CAT NBP2-75298 (NOVUS BIOLOGICALS, Centennial, CO, USA) were used.

Techniques: Clinical Proteomics, Enzyme-linked Immunosorbent Assay

Journal: Cancers

Article Title: New Advances in the Study of CMTM6, a Focus on Its Novel Non-Canonical Cellular Locations, and Functions beyond Its Role as a PD-L1 Stabilizer

doi: 10.3390/cancers16183126

Figure Lengend Snippet: CMTM6 and PD-L1 were found in exosomes from the plasma of the healthy donor group (HD) and the group of patients with cervical cancer (CC). ( a ) CMTM6 is preferentially released in exosomes in both the HD ( n = 10) and CC ( n = 20) groups; data shown as pg/mL of CMTM6. ( b ) PD-L1 was elevated in the exosome-enriched plasma fractions from both the HDs ( n = 6) and CC ( n = 15); data shown as pg/mL. ( c ) Correlation between exosomal CMTM6 and PD-L1 in HD samples ( r = 0.996). ( d ) Correlation between exosomal CMTM6 and PD-L1 in CC samples ( r = 0.8346). ( e ) CMTM6 was found to increase in the exosomal fractions from HDs and CC patients by Western blot. CD63 was used as an exosomal marker. The uncropped blots are shown in . Wilcoxon ranked test was used. * p ≤ 0.05, ** p ≤ 0.01, *** p < 0.005.

Article Snippet: Human PD-L1 ELISA Kit, Lot 356381-001, REF BMS2327 (ThermoFisher, Waltham, MA, USA) and HUMAN CMTM6 ELISA Kit (Colorimetric), CAT NBP2-75298 (NOVUS BIOLOGICALS, Centennial, CO, USA) were used.

Techniques: Clinical Proteomics, Western Blot, Marker

Journal: Cancers

Article Title: New Advances in the Study of CMTM6, a Focus on Its Novel Non-Canonical Cellular Locations, and Functions beyond Its Role as a PD-L1 Stabilizer

doi: 10.3390/cancers16183126

Figure Lengend Snippet: CMTM6 and PD-L1 were found in the lysates of CC-derived cells. ( a ) CMTM6 expression was found to be higher in the total lysate of CaSki cells compared to those of HeLa and SiHa cells, shown as bands corresponding to the approximate molecular weight reported for CMTM6 and its densitometric analysis. ( b ) The bands corresponding to the molecular weight reported for PD-L1 were also found in the three cell lines, coinciding again to show that CaSki cells express the highest levels of this protein; the band pattern and its densitometric analysis are shown. In both cases, β-actin was used as a constitutive protein and loading control. The uncropped blots are shown in . Data are shown as mean ± SD; three independent experiments were performed for each condition. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001, ns: no significance.

Article Snippet: Human PD-L1 ELISA Kit, Lot 356381-001, REF BMS2327 (ThermoFisher, Waltham, MA, USA) and HUMAN CMTM6 ELISA Kit (Colorimetric), CAT NBP2-75298 (NOVUS BIOLOGICALS, Centennial, CO, USA) were used.

Techniques: Derivative Assay, Expressing, Molecular Weight, Control

Journal: Cancers

Article Title: New Advances in the Study of CMTM6, a Focus on Its Novel Non-Canonical Cellular Locations, and Functions beyond Its Role as a PD-L1 Stabilizer

doi: 10.3390/cancers16183126

Figure Lengend Snippet: CMTM6 was found in the cell membrane and intracellularly in cell lines derived from cervical cancer. ( a ) The percentage of CMTM6-positive cells was determined by flow cytometry. CMTM6 was found both intracellularly and associated with the plasma membranes of all cell lines. Interestingly, the highest percentages of CMTM6, both intracellular and on the membrane, were found in CaSki cells. Data are shown as mean ± SD; three independent experiments were performed for each condition. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001, ns: no significance. ( b ) Immunofluorescence staining verified that CMTM6 (AF-594, red stain) is found in different subcellular locations such as intracellular (cytoplasm shown with diamond-tipped arrow) and the plasma membrane (closed arrow). The nuclei (open arrow) are stained with DAPI (blue). Note the intracellular staining in SiHa cells, which obscures the nucleus, and the visible nucleus and membrane staining in CaSki cells. Images taken using the 10× objective (left side) and 30× (right side).

Article Snippet: Human PD-L1 ELISA Kit, Lot 356381-001, REF BMS2327 (ThermoFisher, Waltham, MA, USA) and HUMAN CMTM6 ELISA Kit (Colorimetric), CAT NBP2-75298 (NOVUS BIOLOGICALS, Centennial, CO, USA) were used.

Techniques: Membrane, Derivative Assay, Flow Cytometry, Clinical Proteomics, Immunofluorescence, Staining

Journal: Cancers

Article Title: New Advances in the Study of CMTM6, a Focus on Its Novel Non-Canonical Cellular Locations, and Functions beyond Its Role as a PD-L1 Stabilizer

doi: 10.3390/cancers16183126

Figure Lengend Snippet: CMTM6 released by cell lines derived from cervical cancer. ELISA of culture supernatants to quantify the total levels of supernatant CMTM6. Detectable levels of supernatant CMTM6 were seen in all cell lines; however, HeLa cells were found to release the highest concentration (11,015 pg/mL). Data are shown as mean ± SD; two independent experiments were performed for each condition. * p < 0.05; *** p < 0.001; **** p < 0.0001.

Article Snippet: Human PD-L1 ELISA Kit, Lot 356381-001, REF BMS2327 (ThermoFisher, Waltham, MA, USA) and HUMAN CMTM6 ELISA Kit (Colorimetric), CAT NBP2-75298 (NOVUS BIOLOGICALS, Centennial, CO, USA) were used.

Techniques: Derivative Assay, Enzyme-linked Immunosorbent Assay, Concentration Assay