mouse tnf α paired antibodies  (R&D Systems)

Journal: International Journal for Parasitology: Drugs and Drug Resistance

Article Title: Terpenic compounds possess anthelmintic and immunomodulatory properties with potential for controlling equine cyathostomin infections

doi: 10.1016/j.ijpddr.2026.100642

Figure Lengend Snippet: Scatter plots of the association between TNF-α production and the IC 50 values (LDA and LMA) Scatter plots showing the association between TNF-α production percentage (expressed relative to the control) and the IC 50 values obtained in (left) the Larval Development Assay (LDA) and (right) the Larval Migration Assay (LMA) for six terpene compounds (anethole, cinnamaldehyde, menthol, carvacrol, eugenol and thymol). Each point represents the mean IC 50 and TNFα production for a given compound, and horizontal/vertical bars indicate the corresponding confidence intervals. Lower IC 50 values reflect higher antiparasitic potency.

Article Snippet: The cells were then incubated at +37 °C (5% CO 2 ) for 24 h. After incubation, the concentration of TNF-α in the medium for each condition was quantified by ELISA using mouse TNF-α paired antibodies (R and D Systems DY410).

Techniques: Control, Migration

Journal: International Journal for Parasitology: Drugs and Drug Resistance

Article Title: Terpenic compounds possess anthelmintic and immunomodulatory properties with potential for controlling equine cyathostomin infections

doi: 10.1016/j.ijpddr.2026.100642

Figure Lengend Snippet: Anti-inflammatory activity of carvacrol and cinnamaldehyde on equine PBMC Boxplots showing TNF-α concentrations (ng/mL) measured in equine peripheral blood mononuclear cells (PBMCs) exposed to DMSO (0.05%), LPS (125 ng/mL), the combination of DMSO and LPS, carvacrol (5 μg/mL), cinnamaldehyde (5 μg/mL), the combination of either compound with LPS, and the untreated condition (control). Points represent individual replicates from four independent assays. Asterisks indicate significant differences relative to the corresponding control condition (∗ P = 0.01, ∗∗ P < 0.001).

Article Snippet: The cells were then incubated at +37 °C (5% CO 2 ) for 24 h. After incubation, the concentration of TNF-α in the medium for each condition was quantified by ELISA using mouse TNF-α paired antibodies (R and D Systems DY410).

Techniques: Activity Assay, Control

Journal: Molecular Medicine Reports

Article Title: Veronicastrum sibiricum (L.) Pennell extract alleviates inflammation-induced muscle atrophy through NLRP3 inflammasome regulation and mitochondrial function restoration

doi: 10.3892/mmr.2026.13857

Figure Lengend Snippet: VSE reduces the secretion of proinflammatory cytokines. C2C12 myotubes were pretreated with VSE at 0 (LPS only), 1.25, 2.5 and 5 µg/ml for 3 h and then co-treated with 500 ng/ml LPS for 24 h. Controls received no treatments. (A) mRNA expression levels of the proinflammatory cytokines TNF-α and IL-1β were analyzed using reverse transcription-quantitative PCR. (B) Protein expression levels of the proinflammatory cytokines TNF-α and IL-1β in the supernatant of C2C12 myotube cultures were analyzed using ELISA. All data are presented as the mean ± SD of triplicate results and statistical analysis was carried out using one-way ANOVA with Tukey's post hoc test. #### P<0.0001 vs. control; ***P<0.001, ****P<0.0001 vs. LPS treatment. A450, absorbance at 450 nm; LPS, lipopolysaccharide; VSE, Veronicastrum sibiricum seed extract.

Article Snippet: Supernatants were acquired, and secretion of the proinflammatory cytokines TNF-α (cat. no. MTA00B; R&D systems, Inc.) and IL-1β was analyzed using ELISA kits (cat. no. DY401; R&D Systems, Inc.) according to the manufacturer's guidelines.

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Control

Journal: Bone Research

Article Title: Targeting adipocyte ESRRA alleviates osteoarthritis via interrupting inter-organelle crosstalk of complement C3-CFD-MAC cascade

doi: 10.1038/s41413-026-00527-3

Figure Lengend Snippet: Reduced CFD from Esrra -ablated BMAds interrupts C3-CFD-MAC cascade-mediated mitochondrial dysfunction, senescence and catabolism in chondrocytes. a , b Representative images ( a ) and quantification ( b ) of MAC and pERK1/2 positive staining in cartilage regions of HFHC-fed or 25-month-old Esrra fl/fl and Esrra AKO mice. Scale bar, 100 μm. n = 6 mice. c Diagram depicting primary wild-type chondrocytes treated with the indicated conditioned medium (CM) collected from mBMAds, with or without supplementation of 1 μg/mL mouse recombinant C3 (mC3). d Soluble CFD concentrations from BMAds CM prepared as in ( c ) ( n = 6). e mRNA levels of degradative enzymes Mmp13 , Mmp3 , Mmp9 , Adamts4 , Adamts5 and senescent/proinflammatory markers P16 , P53 , P21 , IL6 in co-cultured primary chondrocytes as in ( c ) ( n = 5). f Protein levels of pERK1/2, ERK1/2, P21, MMP13 and COL2A1 in co-cultured primary chondrocytes as in ( c ). g , h Immunofluorescence staining of MAC, pERK1/2, MitoTracker Red, MitoSOX Red, γH2AX, P21 and Ki67, as well as SA-β-gal staining in co-cultured primary chondrocytes ( g ). Quantitative analysis results are shown in ( h ) ( n = 4). DAPI or Hoechst was used for nuclear counterstaining. Scale bar, 10 μm. i Oxygen Consumption Rate (OCR) were measured in co-cultured primary chondrocytes as in ( c ). Bar chart showing the results of mitochondrial basal and maximal respiratory capacity changes ( n = 9). Data are shown as mean ± SD. Statistical analysis is performed using two-sided unpaired Student’s t -test ( b , d ), two-way ANOVA with post-hoc Turkey’s multiple comparisons test ( e , h , i )

Article Snippet: ELISA assays were performed to detect CFD (R&D Systems #DY5430-05), C3 (Cloud-clone #SEA861Mu), and P1NP (Novus Biologicals #NBP2-76466).

Techniques: Staining, Recombinant, Cell Culture, Immunofluorescence

Journal: Bone Research

Article Title: Targeting adipocyte ESRRA alleviates osteoarthritis via interrupting inter-organelle crosstalk of complement C3-CFD-MAC cascade

doi: 10.1038/s41413-026-00527-3

Figure Lengend Snippet: Inhibition of adipocyte ESRRA/CFD signaling rescues chondrocyte from damage caused by excessive hepatocytes-derived C3 under metabolic stress conditions. a Boxplots depicting C3 gene expression levels in MASLD/MASH patients compared to healthy controls from European ( GSE135251 ) and American ( GSE24807 ) cohorts. Data are represented as box and whiskers with bars representing maximum and minimum values and with median highlighted as a line. b Immunoblot analysis of C3 expression in livers from female mice at various time points following OVX (left) and male mice across different ages (right). c Representative liver images of the livers from HFHC-fed or 25-month-old Esrra fl/fl and Esrra AKO mice as well as corresponding controls. d Protein levels of C3 in the livers as in ( c ). e ELISA analysis of serum C3 concentrations. n = 6 mice per group. f The expression and secretion of C3 protein in primary hepatocytes following treatment with 40 μmol/L cholesterol (Chol) or a mixture of 250 μmol/L oleic acid (OA)/125 μmol/L palmitic acid (PA) for 24 hours (n = 4). g Diagram illustrating the co-culture system involving mBMAds CM, mouse primary hepatocytes and chondrocytes. h Western blot analysis of pERK1/2, ERK1/2, P21, MMP13 and COL2A1 in co-cultured primary chondrocytes as in ( g ). i , j Representative images ( i ) and quantitative analysis ( j ) of MAC, pERK1/2, MitoTracker Red, MitoSOX Red, γH2AX, P21, Ki67 and SA-β-gal staining in chondrocytes as in ( g ) ( n = 4). DAPI or Hoechst was used for counterstaining of nuclei. Scale bar, 10 μm. Data are shown as mean ± SD. Statistical analysis is performed using two-sided unpaired Student’s t -test (right panel of a ), two-way ANOVA with post-hoc Turkey’s multiple comparisons test ( e , j ), one-way ANOVA followed by Bonferroni’s post hoc tests (left panel of a , f )

Article Snippet: ELISA assays were performed to detect CFD (R&D Systems #DY5430-05), C3 (Cloud-clone #SEA861Mu), and P1NP (Novus Biologicals #NBP2-76466).

Techniques: Inhibition, Derivative Assay, Gene Expression, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Co-Culture Assay, Cell Culture, Staining

Journal: Bone Research

Article Title: Targeting adipocyte ESRRA alleviates osteoarthritis via interrupting inter-organelle crosstalk of complement C3-CFD-MAC cascade

doi: 10.1038/s41413-026-00527-3

Figure Lengend Snippet: Suppressing CFD by Danicopan or andrographolide confers protection against cellular senescence and catabolism in human C28/I2 chondrocytes. a Scatter plot showing plasma CFD levels across the lifespan were analyzed using a public proteome dataset (see Method for details). RFU, relative fluorescent unit. b Illustration of the co-culture system involving C28/I2 chondrocytes and conditioned medium (CM) obtained from AP-treated human BMAds (hBMAds), with an additional treatment of 1 μg/mL human recombinant C3 (hC3) and/or 10 mmol/L Danicopan. c Western blot analysis of ESRRA, CFD and Leptin protein levels in human BMSCs-derived BMAds treated with 40 μmol/L AP or DMSO for 2 days. d Western blot analysis of pERK1/2, ERK1/2, P21, MMP13, and COL2A1 protein levels in human C28/I2 chondrocytes as in ( b ). e , f Representative images ( e ) and corresponding quantitative data ( f ) of MAC, pERK1/2, MitoTracker Red, MitoSOX Red, γH2AX, P21, Ki67 and SA-β-gal staining in human C28/I2 chondrocytes as in ( b ) ( n = 4). DAPI or Hoechst was used for nuclear counterstaining. Scale bar, 10 μm. Data are shown as mean ± SD. Statistical analysis is performed using two-way ANOVA with post-hoc Turkey’s multiple comparisons test

Article Snippet: ELISA assays were performed to detect CFD (R&D Systems #DY5430-05), C3 (Cloud-clone #SEA861Mu), and P1NP (Novus Biologicals #NBP2-76466).

Techniques: Clinical Proteomics, Co-Culture Assay, Recombinant, Western Blot, Derivative Assay, Staining

Journal: Bone Research

Article Title: Targeting adipocyte ESRRA alleviates osteoarthritis via interrupting inter-organelle crosstalk of complement C3-CFD-MAC cascade

doi: 10.1038/s41413-026-00527-3

Figure Lengend Snippet: Pharmacological inhibition of ESRRA by andrographolide protects aged mice against spontaneous osteoarthritis progression. a Schematic diagram of the experimental design for pharmacological treatments in mice. 22-month-old male C57BL/6 mice were administered intragastrically (i.g.) with either vehicle or AP (100 mg/kg body weight) five times per week for 3 months. Young controls were 3-month-old male mice. b Representative images of WAT depots and livers, along with WAT weight analysis. c Liver TC and TG levels. d Immunoblots of C3 in liver tissues. e Serum C3 levels. f Immunoblots of bone marrow CFD and Leptin. g Serum CFD levels. h , i Immunofluorescence of PLIN1 staining ( h ) and quantification of PLIN1 + bone marrow adipocytes area ( i ). Scale bar, 100 μm. j Representative SO&FG-stained joint sections and H&E-stained synovial tissue. Scale bar, 100 μm. k , l OARSI score ( k ) and synovitis score ( l ). m Digital microCT images of knee joints. n , Quantitative data of the volume of calcified meniscus and synovial tissue. o , p Immunofluorescence staining for MAC and pERK1/2 in articular cartilage ( o ) and the quantitation ( p ). Scale bar, 100 μm. q – t Immunofluorescence staining ( q , r ) and quantification ( s , t ) of p21, Ki67, MMP13 and COL2A1. Scale bar, 100 μm. u A proposed model illustrates that ESRRA-mediated adipocyte CFD and liver-derived C3 synergistically promote osteoarthritis progression in aging and metabolic disorders. ESRRA transcriptionally upregulates CFD responding to MAT expansion. Together with steatotic liver-derived C3, this triggers excessive alternative complement activation, resulting in MAC deposition on chondrocytes, therefore provoking pERK1/2 activation and mitochondrial dysfunction. Targeting ESRRA alleviates osteoarthritis by interrupting this inter-organ C3-CFD-MAC cascade. For all experiments, n = 6 mice. Data are shown as mean ± SD. Statistical analysis is performed using two-way ANOVA with post-hoc Turkey’s multiple comparisons test

Article Snippet: ELISA assays were performed to detect CFD (R&D Systems #DY5430-05), C3 (Cloud-clone #SEA861Mu), and P1NP (Novus Biologicals #NBP2-76466).

Techniques: Inhibition, Western Blot, Immunofluorescence, Staining, Quantitation Assay, Derivative Assay, Activation Assay

Journal: Bone Research

Article Title: Vitamin K-dependent carboxylation in osteoblasts regulates bone resorption through GAS6 in male mice

doi: 10.1038/s41413-026-00528-2

Figure Lengend Snippet: The γ-carboxylated protein GAS6 is expressed by osteoblasts and activates its receptors on pre-osteoclasts. a Expression analysis by qPCR of genes encoding the known γ-carboxylated proteins. Gas6 is highlighted in red. b Gene expression analysis by qPCR of the TAM receptors Axl , Mertk and Tyro3 in bone marrow derived monocytes (d0: day 0) and in differentiating osteoclast cultures in the presence of RANKL and M-CSF (d2-d6: day 2 to 6; n = 4). c Western blot analysis of the phosphorylation (P) of AKT (S473) and TAM (Y702 in AXL and Y753 in MerTK) in bone marrow derived monocytes (BMMC) serum starved for 3 h and treated with GAS6 (200 ng/mL) for the indicated times. Total AKT, MerTK, and AXL were used as loading controls. d Quantification of P-AKT (S473), P-MerTK (Y753), and P-AXL (Y702) normalized to the amount of total protein ( n = 3). Results represent the mean ± SEM

Article Snippet: GAS6 from serum and osteoblast supernatant were quantified using a mouse GAS6 ELISA (DuoSet ELISA DY986, R&D Systems, Minneapolis, MN) as we previously described.

Techniques: Expressing, Gene Expression, Derivative Assay, Western Blot, Phospho-proteomics

Journal: Bone Research

Article Title: Vitamin K-dependent carboxylation in osteoblasts regulates bone resorption through GAS6 in male mice

doi: 10.1038/s41413-026-00528-2

Figure Lengend Snippet: TAM signaling and γ-carboxylated GAS6 signaling promotes osteoclast formation in culture. Representative TRAP staining (left) and quantification of the TRAP + osteoclast area (right) in WT osteoblast (OB) and bone marrow cell (BM) co-cultures at day 8 in the presence of PGE 2 and VitD 3 , with or without the TAM inhibitors LDC1267 ( a ) or R428 ( b ), at the indicated concentrations. c–f Bone marrow derived monocytes (BMMC) were cultured in the presence of RANKL (20 ng/mL) and M-CSF (10 ng/mL) with or without recombinant γ-carboxylated GAS6 at the indicated concentrations for up to 6 days. c Representative TRAP staining at day 5 and 6 of differentiation. d Quantification of the TRAP + osteoclast area at day 4, 5, and 6 of differentiation. e Quantification of the number of TRAP + multinucleated osteoclasts at day 4, 5, and 6 of differentiation. f Quantification of the number of nuclei per osteoclast at day 5 of differentiation. Results represent the mean ± SEM. One-way ANOVA with Bonferroni’s posttests was used in ( a , b ) and ( d – f ). *** P < 0.001, ** P < 0.01, * P < 0.05, ns: non-significant

Article Snippet: GAS6 from serum and osteoblast supernatant were quantified using a mouse GAS6 ELISA (DuoSet ELISA DY986, R&D Systems, Minneapolis, MN) as we previously described.

Techniques: Staining, Derivative Assay, Cell Culture, Recombinant

Journal: Bone Research

Article Title: Vitamin K-dependent carboxylation in osteoblasts regulates bone resorption through GAS6 in male mice

doi: 10.1038/s41413-026-00528-2

Figure Lengend Snippet: Gamma-carboxylated GAS6 impacts on osteoclast differentiation and fusion. a–d Gene expression analysis by qPCR of osteoclast differentiation markers Acp5 (TRAP), Clcn7 , Ctsk , and Dcstamp . Bone marrow derived monocytes (BMMC) were cultured in the presence of RANKL (20 ng/mL) and M-CSF (10 ng/mL) with or without recombinant γ-carboxylated GAS6 at the indicated concentrations for 2, 4, and 6 days ( n = 4 per condition). e Schematic representation of the assay used to assess the impact of γ-carboxylated GAS6 on pre-osteoclast fusion in culture using a conditionally activated tdTomato (Tom) reporter (created with BioRender). f Representative pictures of live osteoclast cultures at the indicated time and concentration of recombinant γ-carboxylated GAS6. The stars indicate the presence of fusion events (Tom + cells) in presence of GAS6 at Day 4. g Quantification of the number of fusion events per 10 mm 2 at the indicated time of osteoclasts differentiation ( n = 16 fields per condition). Results represent the mean ± SEM. One-way ANOVA with Bonferroni’s posttests was used in ( a – d ) and ( g ). *** P < 0.001, ** P < 0.01

Article Snippet: GAS6 from serum and osteoblast supernatant were quantified using a mouse GAS6 ELISA (DuoSet ELISA DY986, R&D Systems, Minneapolis, MN) as we previously described.

Techniques: Gene Expression, Derivative Assay, Cell Culture, Recombinant, Concentration Assay

Journal: Bone Research

Article Title: Vitamin K-dependent carboxylation in osteoblasts regulates bone resorption through GAS6 in male mice

doi: 10.1038/s41413-026-00528-2

Figure Lengend Snippet: GAS6 promotes osteoclast formation and bone resorption in vivo. a–j Six-month-old WT (non-transgenic littermates) and ApoE-Gas6 Tg male mice were analyzed. GAS6 concentration in the serum ( a ) and bone marrow cavity ( b ) ( n = 4). c Representative pictures of sections from lumbar vertebrae stained with von Kossa and van Gieson. d Quantification of trabecular bone volume over tissue volume (BV/TV) from the L4 and L5 lumbar vertebrae sections ( n = 15–17). e–h μCT analysis of the distal femur trabecular bone ( n = 12). e Representative μCT images. f Quantification of trabecular bone volume (BV/TV). g Quantification of trabecular bone surface density (BS/TV). h Trabecular bone µCT derived data. Tb.Sp., Tb.N., and Tb.Th., trabecular spacing, number, and thickness, respectively; Conn.Dn., connectivity density. i–j Bone histomorphometry analysis of lumbar vertebrae in six-month-old WT and ApoE-Gas6 Tg male ( n = 19–20). i Representative pictures of TRAP staining. j Number of osteoclasts per bone perimeter (N.Oc/B.Pm) and osteoclast surface over bone surface (Oc.S/BS). Results represent the mean ± SEM. Unpaired, 2-tailed Student’s t test was used in ( a , b , d , f – h and j ). *** P < 0.001, ** P < 0.01, * P < 0.05, ns: non-significant

Article Snippet: GAS6 from serum and osteoblast supernatant were quantified using a mouse GAS6 ELISA (DuoSet ELISA DY986, R&D Systems, Minneapolis, MN) as we previously described.

Techniques: In Vivo, Transgenic Assay, Concentration Assay, Staining, Derivative Assay

Journal: Bone Research

Article Title: Vitamin K-dependent carboxylation in osteoblasts regulates bone resorption through GAS6 in male mice

doi: 10.1038/s41413-026-00528-2

Figure Lengend Snippet: Increased bone mass in male mice lacking γ-carboxylation in osteoblasts. a Gene expression analysis by qPCR of Ggcx and Vkorc1 in bone marrow derived monocytes (BMMC), osteoclasts (OCL), proliferating pre-osteoblasts (pre-OB), and mineralized osteoblast (OB) cultures ( n = 3). b Protein expression in liver (Liv) and bone cells by Western blot. GGCX was analyzed on a 7.5% SDS Tris Glycine gel using 20 μg of extracts, while VKORC1 was resolved on a 10% SDS Tris Tricine gel using 10 μg of extracts. c–g Six-month-old Ggcx ff and Ggcx ff ;OCN-Cre male mice were analyzed ( n = 7–10). c Representative pictures of sections from lumbar vertebrae stained with von Kossa and van Gieson. d Quantification of trabecular bone volume over tissue volume (BV/TV) from the L4 and L5 lumbar vertebrae sections. e Representative μCT images of the distal femur trabecular bones. f Quantification of trabecular bone volume (BV/TV) and trabecular bone surface density (BS/TV) from the μCT data. g Trabecular bone µCT derived data. Tb.Sp., Tb.N., and Tb.Th., trabecular spacing, number, and thickness respectively; Conn.Dn., connectivity density. Unpaired, 2-tailed Student’s t test was used in ( d ), ( f ), and ( g ). *** P < 0.001, ** P < 0.01, * P < 0.05, ns: non-significant

Article Snippet: GAS6 from serum and osteoblast supernatant were quantified using a mouse GAS6 ELISA (DuoSet ELISA DY986, R&D Systems, Minneapolis, MN) as we previously described.

Techniques: Gene Expression, Derivative Assay, Expressing, Western Blot, Staining

Journal: Bone Research

Article Title: Vitamin K-dependent carboxylation in osteoblasts regulates bone resorption through GAS6 in male mice

doi: 10.1038/s41413-026-00528-2

Figure Lengend Snippet: Reduced osteoclast number and surface in Ggcx ff ;OCN-Cre male mice. a–h Bone histomorphometry analysis of lumbar vertebrae in six-month-old Ggcx ff and Ggcx ff ;OCN-Cre male mice ( n = 6–10). a Representative pictures of calcein double labeling and toluidine blue staining. b Mineral apposition rate (MAR). c Bone formation rate over bone surface (BFR/BS). d Number of osteoblasts per bone perimeter (N.Ob/B.Pm). e Osteoblast surface over bone surface (Ob.S/BS). f Representative pictures of TRAP staining. g Number of osteoclasts per bone perimeter (N.Oc/B.Pm). h Osteoclast surface over bone surface (Oc.S/BS). i Fasting serum CTx levels ( n = 12–17). Results represent the mean ± SEM. Unpaired, 2-tailed Student’s t test was used in ( b – e ) and ( g – i ). ** P < 0.01, * P < 0.05, ns: non-significant

Article Snippet: GAS6 from serum and osteoblast supernatant were quantified using a mouse GAS6 ELISA (DuoSet ELISA DY986, R&D Systems, Minneapolis, MN) as we previously described.

Techniques: Labeling, Staining

Journal: Bone Research

Article Title: Vitamin K-dependent carboxylation in osteoblasts regulates bone resorption through GAS6 in male mice

doi: 10.1038/s41413-026-00528-2

Figure Lengend Snippet: Ggcx inactivation impairs the ability of osteoblasts to support osteoclastogenesis ex vivo. a Representative TRAP staining of osteoblasts (OB) and bone marrow cells (BM) co-cultures at day 8 in the presence of prostaglandin E 2 (PGE 2 ; 10 –6 mol/L) and 1,25 vitamin D 3 (VitD 3 ; 10 –8 mol/L). Ggcx ff osteoblasts were transduced with either Ad-GFP (control) or Ad-Cre (knockout) before the addition of the WT bone marrow cells. b Quantification of the number of TRAP + osteoclasts per well (Nb of OCL/well) ( n = 3). c Representative TRAP staining of osteoblasts and bone marrow cells co-cultures at day 8. Control ( Ocn + / + ) or osteocalcin-deficient ( Ocn -/- ) osteoblasts were cultured with WT bone marrow cells. d Quantification of the number of TRAP + osteoclasts per well (Nb of OCL/well) ( n = 3). e Gene expression analysis by qPCR in Ggcx ff + Ad-GFP and Ggcx ff + Ad-Cre osteoblasts cultured in presence (+) or absence (–) of PGE 2 and VitD 3 for 6 days. Results represent the mean ± SEM. Unpaired, 2-tailed Student’s t test was used in ( b ) and ( d ). Two-way ANOVA with Bonferroni’s posttests was used in ( e ). *** P < 0.001, ** P < 0.01, * P < 0.05, ns: non-significant

Article Snippet: GAS6 from serum and osteoblast supernatant were quantified using a mouse GAS6 ELISA (DuoSet ELISA DY986, R&D Systems, Minneapolis, MN) as we previously described.

Techniques: Ex Vivo, Staining, Transduction, Control, Knock-Out, Cell Culture, Gene Expression

Journal: Bone Research

Article Title: Vitamin K-dependent carboxylation in osteoblasts regulates bone resorption through GAS6 in male mice

doi: 10.1038/s41413-026-00528-2

Figure Lengend Snippet: The γ-carboxylated protein GAS6 is expressed by osteoblasts and activates its receptors on pre-osteoclasts. a Expression analysis by qPCR of genes encoding the known γ-carboxylated proteins. Gas6 is highlighted in red. b Gene expression analysis by qPCR of the TAM receptors Axl , Mertk and Tyro3 in bone marrow derived monocytes (d0: day 0) and in differentiating osteoclast cultures in the presence of RANKL and M-CSF (d2-d6: day 2 to 6; n = 4). c Western blot analysis of the phosphorylation (P) of AKT (S473) and TAM (Y702 in AXL and Y753 in MerTK) in bone marrow derived monocytes (BMMC) serum starved for 3 h and treated with GAS6 (200 ng/mL) for the indicated times. Total AKT, MerTK, and AXL were used as loading controls. d Quantification of P-AKT (S473), P-MerTK (Y753), and P-AXL (Y702) normalized to the amount of total protein ( n = 3). Results represent the mean ± SEM

Article Snippet: GAS6 from serum and osteoblast supernatant were quantified using a mouse GAS6 ELISA (DuoSet ELISA DY986, R&D Systems, Minneapolis, MN) as we previously described.

Techniques: Expressing, Gene Expression, Derivative Assay, Western Blot, Phospho-proteomics

Journal: Bone Research

Article Title: Vitamin K-dependent carboxylation in osteoblasts regulates bone resorption through GAS6 in male mice

doi: 10.1038/s41413-026-00528-2

Figure Lengend Snippet: TAM signaling and γ-carboxylated GAS6 signaling promotes osteoclast formation in culture. Representative TRAP staining (left) and quantification of the TRAP + osteoclast area (right) in WT osteoblast (OB) and bone marrow cell (BM) co-cultures at day 8 in the presence of PGE 2 and VitD 3 , with or without the TAM inhibitors LDC1267 ( a ) or R428 ( b ), at the indicated concentrations. c–f Bone marrow derived monocytes (BMMC) were cultured in the presence of RANKL (20 ng/mL) and M-CSF (10 ng/mL) with or without recombinant γ-carboxylated GAS6 at the indicated concentrations for up to 6 days. c Representative TRAP staining at day 5 and 6 of differentiation. d Quantification of the TRAP + osteoclast area at day 4, 5, and 6 of differentiation. e Quantification of the number of TRAP + multinucleated osteoclasts at day 4, 5, and 6 of differentiation. f Quantification of the number of nuclei per osteoclast at day 5 of differentiation. Results represent the mean ± SEM. One-way ANOVA with Bonferroni’s posttests was used in ( a , b ) and ( d – f ). *** P < 0.001, ** P < 0.01, * P < 0.05, ns: non-significant

Article Snippet: GAS6 from serum and osteoblast supernatant were quantified using a mouse GAS6 ELISA (DuoSet ELISA DY986, R&D Systems, Minneapolis, MN) as we previously described.

Techniques: Staining, Derivative Assay, Cell Culture, Recombinant

Journal: bioRxiv

Article Title: Increasing plasma bile salt levels with Bulevirtide alleviates DSS-induced colitis and LPS-induced inflammation

doi: 10.64898/2026.04.21.719641

Figure Lengend Snippet: (A) Experimental schematic; BMDMs were treated with TCDC (100 µM) for 1 hour, followed by LPS (10ng/mL) for 3 hours. To activate the inflammasome, cells were also treated with ATP (1mM, 30 minutes before harvesting). (B) Gene expression of the cytokines TNF, MCP1, IL-1β, IL-10 and IL-12 in BMDMs. (C) TNF and (D) IL10 concentration in medium of BMDMs was measured by ELISA. (E) Protein levels of IL-1β and caspase-1 were measured in whole cell lysates of BMDMs. (F) IL-1β and (G) IL-18 concentrations in medium of BMDMs were measured by ELISA. (H) TCA uptake of BMDMs, in comparison to U2OS parental and U2OS-NTCP cells. (I) Caspase 3/7 activity was assessed as a marker of apoptotic activity in BMDMs incubated with the indicated concentrations of TCDC. n=3-6 per group. Experiments were performed 3 times and representative data are shown. Data are means ± SD. Significance was assessed with Mann-Whitney U test. * p <0.05.

Article Snippet: IL-10, IL-1β and TNF concentrations were measured in BMDM supernatant by ELISA (cat. No. DY401, DY417, DY410, R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s instruction.

Techniques: Gene Expression, Concentration Assay, Enzyme-linked Immunosorbent Assay, Comparison, Activity Assay, Marker, Incubation, MANN-WHITNEY

Journal: bioRxiv

Article Title: Increasing plasma bile salt levels with Bulevirtide alleviates DSS-induced colitis and LPS-induced inflammation

doi: 10.64898/2026.04.21.719641

Figure Lengend Snippet: (A) Schematic experimental overview; Slco1a/1b -/- FVB mice were injected with vehicle or Bulevirtide 2.5 µg/g, followed by LPS injection at 20 µg/g. (B) Total bile salt concentration and (C) concentration of individual bile salt species measured by HPLC. Plasma concentration of the cytokines TNF (D), IL-10 (E), IL-1β (F), IL-12p70 (G), IL-6 (H) and MCP1 (I). IL-1β was measured by ELISA, whereas other cytokines were measured by cytometric bead array. n=8 per group. Data are means ± SD. Significance was assessed with Mann-Whitney U test. * p <0.05.

Article Snippet: IL-10, IL-1β and TNF concentrations were measured in BMDM supernatant by ELISA (cat. No. DY401, DY417, DY410, R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s instruction.

Techniques: Injection, Concentration Assay, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY