recombinant human gp130 fc chimera protein  (R&D Systems)

Journal: Journal of Medicinal Chemistry

Article Title: Development of the First Small-Molecule Inhibitor Targeting Oncostatin M for Treatment of Breast Cancer

doi: 10.1021/acs.jmedchem.4c03233

Figure Lengend Snippet: OSM structure and signaling. (A) In order to form a complete signaling complex, OSM first binds to gp130, which then allows for the recruitment of LIFRβ or OSMRβ. Afterward, several signaling cascades are activatedmainly the JAK/STAT3, PI3K/AKT, MAPK/ERK, and JNK pathways (not shown). (B) OSM is a four α-helical bundle protein with two distinct binding pockets for ligand–receptor interaction. Site II is responsible for gp130 binding and Site III is utilized for interaction with OSMRβ.

Article Snippet: Protein used: gp130 (R&D Systems, cat. #671-GP) and OSMRβ (R&D Systems, cat. # 4389-OR).

Techniques: Binding Assay

Journal: International Journal of Molecular Sciences

Article Title: Nonspecific Inhibition of IL6 Family Cytokine Signalling by Soluble gp130

doi: 10.3390/ijms25031363

Figure Lengend Snippet: sgp130Fc inhibits IL6, IL11, OSM, and CT1-induced STAT3 activation. ( A ) ELISA of IL6R, IL11RA, LIFR, and OSMR in the conditioned supernatant of A549, primary human hepatocytes (HEP), and primary human hepatic stellate cells (HSC) 17 h after plating. Data are shown as box-and-whisker plots with median (middle line), 25th–75th percentiles (box) and min-max percentiles (whiskers). Western blots of p -STAT3 and STAT3 in CNTF, CT1, IL6, IL11, LIF (5 ng/mL)-stimulated ( B ) A549, ( C ) HEP, ( D ) HSC (15 min) and ( B – D ) immunoblots of p -STAT3, STAT3, GAPDH in OSM (5 ng/mL)-stimulated A549, HEP, HSC (15 min) in the presence or absence of sgp130Fc (5 µg/mL). ( A – D ) n = 4 biological replicates. BL: baseline. n.d. not detected.

Article Snippet: Recombinant human (rh)CNTF (257-NT, R&D Systems, Minneapolis, MN, USA), rhCT1 (612-CD, R&D Systems, Minneapolis, MN, USA), rhIL6 (206-IL, R&D Systems, Minneapolis, MN, USA), rhIL11 (Z03108, Genscript, Minneapolis, MN, USA), rhLIF (7734-LF, R&D Systems, Minneapolis, MN, USA), rhOSM (PHC5015, ThermoFisher Scientific, San Francisco, CA, USA), sgp130Fc (671-GP, R&D Systems, Minneapolis, MN, USA).

Techniques: Activation Assay, Enzyme-linked Immunosorbent Assay, Whisker Assay, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Nonspecific Inhibition of IL6 Family Cytokine Signalling by Soluble gp130

doi: 10.3390/ijms25031363

Figure Lengend Snippet: Dose-dependent inhibition of IL6 and OSM cis -signalling by sgp130Fc. ( A ) Western blots of STAT3 activation status ( p -STAT3/STAT3) in A549 and HEP in the following conditions: without stimulus (BL), IL6 + PBS, or IL6 + sgp130Fc in the presence of either DMSO or GW280264X. ( B ) ELISA of IL6R in the supernatant of A549 after 17 h incubation with either DMSO or GW280264X. Data are shown as box-and-whisker plots with median (middle line), 25th–75th percentiles (box) and min-max percentiles (whiskers); 2-tailed Student’s t -test. ( C ) Western blots and ( D ) densitometry analyses of p -STAT3/STAT3 in GW280264X-treated A549 following stimulation with IL6 in the presence of different concentrations of sgp130Fc. ( E ) Western blots of p -STAT3, STAT3, and GAPDH, and ( F ) densitometry analyses of p -STAT3/STAT3 in OSM-stimulated A549 in the presence of increasing concentrations of sgp130Fc. ( G ) Western blots and ( H ) densitometry analyses of p -STAT3/STAT3 in IL6-stimulated HEP and HSC in the presence of increasing concentrations of sgp130Fc. ( D , F , H ) Data are shown as mean ± s.d. ( A – H ) n = 4 biological replicates; DMSO (0.1%), GW280264X (1 µM), IL6 (5 ng/mL), OSM (5 ng/mL), sgp130Fc (5 µg/mL, unless otherwise specified). BL: baseline. n.d. not detected.

Article Snippet: Recombinant human (rh)CNTF (257-NT, R&D Systems, Minneapolis, MN, USA), rhCT1 (612-CD, R&D Systems, Minneapolis, MN, USA), rhIL6 (206-IL, R&D Systems, Minneapolis, MN, USA), rhIL11 (Z03108, Genscript, Minneapolis, MN, USA), rhLIF (7734-LF, R&D Systems, Minneapolis, MN, USA), rhOSM (PHC5015, ThermoFisher Scientific, San Francisco, CA, USA), sgp130Fc (671-GP, R&D Systems, Minneapolis, MN, USA).

Techniques: Inhibition, Western Blot, Activation Assay, Enzyme-linked Immunosorbent Assay, Incubation, Whisker Assay

Journal: International Journal of Molecular Sciences

Article Title: Nonspecific Inhibition of IL6 Family Cytokine Signalling by Soluble gp130

doi: 10.3390/ijms25031363

Figure Lengend Snippet: Examples of pre-clinical and clinical studies where sgp130Fc was given at doses that exceeded its half-maximal inhibitory (IC 50 ) concentrations for IL6 or OSM cis -signalling. Dose per mouse reports published values or is estimated based on an average adult mouse mass of 25 g. Estimated maximum concentrations for mouse studies were calculated based on the assumption that sgp130Fc is confined and equally distributed across the extracellular fluid spaces (interstitial fluid and plasma), which represents ~20% of the body weight of an adult mouse [ 20 ]. i.p.: intraperitoneal; i.v.: intravenous, d: day.

Article Snippet: Recombinant human (rh)CNTF (257-NT, R&D Systems, Minneapolis, MN, USA), rhCT1 (612-CD, R&D Systems, Minneapolis, MN, USA), rhIL6 (206-IL, R&D Systems, Minneapolis, MN, USA), rhIL11 (Z03108, Genscript, Minneapolis, MN, USA), rhLIF (7734-LF, R&D Systems, Minneapolis, MN, USA), rhOSM (PHC5015, ThermoFisher Scientific, San Francisco, CA, USA), sgp130Fc (671-GP, R&D Systems, Minneapolis, MN, USA).

Techniques: Clinical Proteomics, Concentration Assay

Journal: International Journal of Molecular Sciences

Article Title: Nonspecific Inhibition of IL6 Family Cytokine Signalling by Soluble gp130

doi: 10.3390/ijms25031363

Figure Lengend Snippet: Comparison of the inhibitory effects of sgp130Fc versus a gp130 neutralising antibody on IL6 or OSM cis -signalling.( A ) Western blots of p -STAT3, STAT3 (for IL6 and OSM), and GAPDH (for OSM) and ( B ) densitometry analyses of p -STAT3/STAT3 in GW280264X-treated A549 following stimulation with IL6, or OSM in the presence of either IgG, sgp130Fc, or a neutralising antibody against gp130 (clone B-R3). ( C ) Levels of IL6R, and OSMR protein in the supernatant from GW280264X-treated A549 as quantified by ELISA. ( D ) Schematic showing our proposed interpretation of the comparative effects between low versus high concentrations of sgp130Fc on the inhibition of gp130-mediated cis - and/or trans- signalling. ( A – C ) n = 4 biological replicates; B-R3 (5 µg/mL), GW280264X (1 µM), IgG (5 µg/mL), IL6 (5 ng/mL), IL11 (5 ng/mL), OSM (5 ng/mL), sgp130Fc (100 µg/mL). ( B , C ) Data are shown as box-and-whisker plots with median (middle line), 25th–75th percentiles (box) and min-max percentiles (whiskers). ( B ) One-way ANOVA with Tukey’s correction. BL: baseline; FC: fold change. n.d. not detected.

Article Snippet: Recombinant human (rh)CNTF (257-NT, R&D Systems, Minneapolis, MN, USA), rhCT1 (612-CD, R&D Systems, Minneapolis, MN, USA), rhIL6 (206-IL, R&D Systems, Minneapolis, MN, USA), rhIL11 (Z03108, Genscript, Minneapolis, MN, USA), rhLIF (7734-LF, R&D Systems, Minneapolis, MN, USA), rhOSM (PHC5015, ThermoFisher Scientific, San Francisco, CA, USA), sgp130Fc (671-GP, R&D Systems, Minneapolis, MN, USA).

Techniques: Comparison, Western Blot, Enzyme-linked Immunosorbent Assay, Inhibition, Whisker Assay