Journal: International Journal of Experimental Pathology

Article Title: Soluble gp130 promotes intestinal epithelial hyperplasia during reovirus infection

doi: 10.1111/j.1365-2613.2010.00710.x

Figure Lengend Snippet: T3D reovirus gene expression at days 2, 4, and 6 post-infection in small intestinal tissues from non-treated and sgp130-treated infected mice. Data are mean values ± SEM; statistically-significant difference (*P< 0.05). No viral RNA was detected in non-infected mice (data not shown).

Article Snippet: Virus, animal infection, and sgp130 treatment T3D reovirus was purchased from American Type Culture Collection (Manassa, VA, USA).

Techniques: Gene Expression, Infection

Journal: International Journal of Experimental Pathology

Article Title: Soluble gp130 promotes intestinal epithelial hyperplasia during reovirus infection

doi: 10.1111/j.1365-2613.2010.00710.x

Figure Lengend Snippet: (a) Villi length and (b) crypt depths at days 2, 4, and 6 post-infection in small intestinal tissues from non-treated and sgp130-treated infected mice. Panels C-E are representative tissue sections from reovirus-infected mice without (d) and with (e) sgp130 treatment at the day 6 post-infection time point, and (c) a tissue section of a normal non-infected mouse. Bars are 100 μm. Data are mean values ± SEM; treatment with sgp130 resulted in statistically-significant less (*P< 0.05; **P< 0.01) villus and crypt shortening at all time points. Statistically shorter (**P< 0.05) crypt length compared to non-infected mice.

Article Snippet: Virus, animal infection, and sgp130 treatment T3D reovirus was purchased from American Type Culture Collection (Manassa, VA, USA).

Techniques: Infection

Journal: International Journal of Experimental Pathology

Article Title: Soluble gp130 promotes intestinal epithelial hyperplasia during reovirus infection

doi: 10.1111/j.1365-2613.2010.00710.x

Figure Lengend Snippet: (a) Treatment of reovirus-infected mice with sgp130 resulted in statistically-significant (*P< 0.05; **P< 0.01) more cells undergoing mitosis at days 2, 4, and 6 post-infection compared to non-treated infected mice. Statistically fewer (*,**P< 0.01) crypt mitotic figures compared to non-infected mice. (b) Example of Cdc47 staining. (c) Crypts of reovirus-infected mice treated with sgp130 had statistically-significant (P < 0.01) more Cdc47+ cells compared to infected non-treated mice. Data are mean values ± SEM.

Article Snippet: Virus, animal infection, and sgp130 treatment T3D reovirus was purchased from American Type Culture Collection (Manassa, VA, USA).

Techniques: Infection, Staining

Journal: Journal of Periodontal & Implant Science

Article Title: Interleukin-6 regulates human ODAM gene expression in gingival epithelial cells

doi: 10.5051/jpis.2402980149

Figure Lengend Snippet: Exposure to IL-6 (10 ng/mL, 12 hours) induced increases in ODAM mRNA levels in Ca9-22 cells (A), HSY cells (B), and Sa3 cells (C), which were inhibited by SC144 (1 μM). ODAM and GAPDH mRNA levels were analyzed using real-time PCR. The experiments were performed in triplicate. Quantitative analyses of the data sets are presented with standard deviations. ODAM: odontogenic ameloblast-associated protein, IL-6: interleukin-6, GAPDH: glyceraldehyde 3-phosphate dehydrogenase, PCR: polymerase chain reaction. Significant differences from the control are indicated by ** P <0.01.

Article Snippet: Twelve hours after transfection, the cells were switched to a serum-free medium for 12 hours and then treated with one of the following inhibitors for 30 minutes: 5 μM protein kinase C inhibitor H7 (Seikagaku Corporation, Tokyo, Japan), 100 nM protein kinase A inhibitor KT5720 (Sigma-Aldrich), 5 μM mitogen-activated protein kinase kinase 1/2 (MEK1/2) inhibitor U0126 (Promega), 10 μM phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002 (Calbiochem, San Diego, CA, USA), 100 nM NF-κB inhibitor triptolide (Tocris Bioscience, Bristol, UK), 1 μM glycoprotein 130 (gp130) inhibitor SC144 (MedChemExpress, Monmouth Junction, NJ, USA), 10 μM STAT1 inhibitor fludarabine (NSC 118218; Selleck Chemicals, Houston, TX, USA), or 10 μM STAT3 inhibitor stattic (Axon Medchem, Reston, VA, USA).

Techniques: Real-time Polymerase Chain Reaction, Polymerase Chain Reaction, Control

Journal: Journal of Periodontal & Implant Science

Article Title: Interleukin-6 regulates human ODAM gene expression in gingival epithelial cells

doi: 10.5051/jpis.2402980149

Figure Lengend Snippet: Protein levels were assessed using Western blotting following stimulation with IL-6 (10 ng/mL) for durations of 10 minutes, 30 minutes, 1 hour, 2 hours, 3 hours, 6 hours, 12 hours, and 24 hours in Ca9-22 cells. Densitometric analyses of the Western blot results for ODAM, p-STAT3, STAT3, p-p65, p65, p-IKKα/β, IKKβ, and IKKα are presented. The intensity of the α-tubulin band was normalized to 1, and graphs were generated to illustrate the relative intensities of the other protein bands. IL-6: interleukin-6, ODAM: odontogenic ameloblast-associated protein, STAT3: signal transducer and activator of transcription 3, p-STAT3: phosphorylated STAT3, p65: nuclear factor-kappa B p65, p-p65: phosphorylated p65, IKKα: IκB kinase α, IKKβ: IκB kinase β, p-IKKα/β: phosphorylated IκB kinase α/β. The data represent the mean ± standard deviation from 3 independent experiments, with * P <0.05 and ** P <0.01 indicating statistical significance.

Article Snippet: Twelve hours after transfection, the cells were switched to a serum-free medium for 12 hours and then treated with one of the following inhibitors for 30 minutes: 5 μM protein kinase C inhibitor H7 (Seikagaku Corporation, Tokyo, Japan), 100 nM protein kinase A inhibitor KT5720 (Sigma-Aldrich), 5 μM mitogen-activated protein kinase kinase 1/2 (MEK1/2) inhibitor U0126 (Promega), 10 μM phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002 (Calbiochem, San Diego, CA, USA), 100 nM NF-κB inhibitor triptolide (Tocris Bioscience, Bristol, UK), 1 μM glycoprotein 130 (gp130) inhibitor SC144 (MedChemExpress, Monmouth Junction, NJ, USA), 10 μM STAT1 inhibitor fludarabine (NSC 118218; Selleck Chemicals, Houston, TX, USA), or 10 μM STAT3 inhibitor stattic (Axon Medchem, Reston, VA, USA).

Techniques: Western Blot, Generated, Standard Deviation

Journal: Journal of Periodontal & Implant Science

Article Title: Interleukin-6 regulates human ODAM gene expression in gingival epithelial cells

doi: 10.5051/jpis.2402980149

Figure Lengend Snippet: (A) LUC activity of −85ODAM, −116ODAM, −174ODAM, −200ODAM, −300ODAM, −330ODAM, −480ODAM, −700ODAM, and −950ODAM constructs were increased by IL-6 (10 ng/mL) after 12 hours of treatment in Ca9-22 cells. Transcriptional activity was determined from 4 separate transfections, and the combined values are expressed with SDs. (B) Effects of kinase inhibitors on the LUC activity of −480ODAM, stimulated by IL-6. Transient transfection analyses of −480ODAM in Ca9-22 cells, with or without IL-6 (10 ng/mL) treatment, were conducted alongside the effects of inhibitors of PKC (H7, 5 μM), PKA (KT5720, 100 nM), tyrosine kinase (HA, 1 μM), MEK1/2 (U0126, 5 μM), PI3K (LY294002, 10 μM), NF-κB (triptolide, 100 nM), STAT3 (stattic, 10 µM), STAT1 (fludarabine, 10 µM), and gp130 (SC144, 1 μM). Transcriptional activity was determined from 3 separate transfections, with the combined values expressed with SDs. ODAM: odontogenic ameloblast-associated protein, IL-6: interleukin-6, LUC: luciferase, SD: standard deviation, PKC: protein kinase C, PKA: protein kinase A, MEK1/2: mitogen-activated protein kinase kinase 1/2, PI3K: phosphatidylinositol 3-kinase, NF-κB: nuclear factor-kappa B, STAT: signal transducer and activator of transcription, gp130: glycoprotein 130. Significant differences from the control are indicated by ** P <0.01.

Article Snippet: Twelve hours after transfection, the cells were switched to a serum-free medium for 12 hours and then treated with one of the following inhibitors for 30 minutes: 5 μM protein kinase C inhibitor H7 (Seikagaku Corporation, Tokyo, Japan), 100 nM protein kinase A inhibitor KT5720 (Sigma-Aldrich), 5 μM mitogen-activated protein kinase kinase 1/2 (MEK1/2) inhibitor U0126 (Promega), 10 μM phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002 (Calbiochem, San Diego, CA, USA), 100 nM NF-κB inhibitor triptolide (Tocris Bioscience, Bristol, UK), 1 μM glycoprotein 130 (gp130) inhibitor SC144 (MedChemExpress, Monmouth Junction, NJ, USA), 10 μM STAT1 inhibitor fludarabine (NSC 118218; Selleck Chemicals, Houston, TX, USA), or 10 μM STAT3 inhibitor stattic (Axon Medchem, Reston, VA, USA).

Techniques: Activity Assay, Construct, Transfection, Luciferase, Standard Deviation, Control

Journal: Journal of Periodontal & Implant Science

Article Title: Interleukin-6 regulates human ODAM gene expression in gingival epithelial cells

doi: 10.5051/jpis.2402980149

Figure Lengend Snippet: Supershift gel shift assays were performed using anti-YY1, C/EBPβ, GATA, STAT3, and p-STAT3 antibodies for each reaction. The DNA-protein complexes were then separated by electrophoresis on a 6% polyacrylamide gel and visualized using the ChemiDoc Touch MP Imaging System. YY1: yin yang 1, C/EBP: CCAAT/enhancer-binding protein, GATA: GATA binding protein, STAT3: signal transducer and activator of transcription 3, p-STAT3: phosphorylated STAT3.

Article Snippet: Twelve hours after transfection, the cells were switched to a serum-free medium for 12 hours and then treated with one of the following inhibitors for 30 minutes: 5 μM protein kinase C inhibitor H7 (Seikagaku Corporation, Tokyo, Japan), 100 nM protein kinase A inhibitor KT5720 (Sigma-Aldrich), 5 μM mitogen-activated protein kinase kinase 1/2 (MEK1/2) inhibitor U0126 (Promega), 10 μM phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002 (Calbiochem, San Diego, CA, USA), 100 nM NF-κB inhibitor triptolide (Tocris Bioscience, Bristol, UK), 1 μM glycoprotein 130 (gp130) inhibitor SC144 (MedChemExpress, Monmouth Junction, NJ, USA), 10 μM STAT1 inhibitor fludarabine (NSC 118218; Selleck Chemicals, Houston, TX, USA), or 10 μM STAT3 inhibitor stattic (Axon Medchem, Reston, VA, USA).

Techniques: Gel Shift, Electrophoresis, Imaging, Binding Assay

Journal: Journal of Periodontal & Implant Science

Article Title: Interleukin-6 regulates human ODAM gene expression in gingival epithelial cells

doi: 10.5051/jpis.2402980149

Figure Lengend Snippet: (A) PCR bands corresponding to DNA-protein complexes immunoprecipitated with antibodies indicated that YY1, C/EBPβ, GATA, STAT3, and p-STAT3 interact with chromatin fragments containing the YY1, C/EBP, GATA, and GATE1-3 sequences. These interactions were increased in Ca9-22 cells following stimulation with IL-6 (10 ng/mL). Input DNA served as a control in the PCR analysis. (B) ChIP analyses were conducted to assess the binding of transcription factors to YY1, C/EBP, GATA, and GATE1-3 in the presence of IL-6 and various kinase inhibitors. Treatment with HA, U0126, LY294002, and triptolide almost completely abolished, and KT5720 partially inhibited, the IL-6-induced (10 ng/mL, 12 hours) binding of YY1, C/EBPβ, and GATA transcription factors to their respective DNA sequences. Additionally, treatment with stattic almost completely abolished, while fludarabine did not inhibit, the IL-6-induced binding of STAT3 transcription factors to GATE1-3. ChIP: chromatin immunoprecipitation, YY1: yin yang 1, C/EBP: CCAAT/enhancer-binding protein, GATA: GATA binding protein, GATE1-3: interferon-γ activated transcriptional element sites 1-3, ODAM: odontogenic ameloblast-associated protein, PCR: polymerase chain reaction, STAT3: signal transducer and activator of transcription 3, p-STAT3: phosphorylated STAT3, IL-6: interleukin-6.

Article Snippet: Twelve hours after transfection, the cells were switched to a serum-free medium for 12 hours and then treated with one of the following inhibitors for 30 minutes: 5 μM protein kinase C inhibitor H7 (Seikagaku Corporation, Tokyo, Japan), 100 nM protein kinase A inhibitor KT5720 (Sigma-Aldrich), 5 μM mitogen-activated protein kinase kinase 1/2 (MEK1/2) inhibitor U0126 (Promega), 10 μM phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002 (Calbiochem, San Diego, CA, USA), 100 nM NF-κB inhibitor triptolide (Tocris Bioscience, Bristol, UK), 1 μM glycoprotein 130 (gp130) inhibitor SC144 (MedChemExpress, Monmouth Junction, NJ, USA), 10 μM STAT1 inhibitor fludarabine (NSC 118218; Selleck Chemicals, Houston, TX, USA), or 10 μM STAT3 inhibitor stattic (Axon Medchem, Reston, VA, USA).

Techniques: Immunoprecipitation, Control, Binding Assay, Chromatin Immunoprecipitation, Polymerase Chain Reaction

Journal: Journal of Periodontal & Implant Science

Article Title: Interleukin-6 regulates human ODAM gene expression in gingival epithelial cells

doi: 10.5051/jpis.2402980149

Figure Lengend Snippet: IL-6–regulated ODAM gene expression is supported by protein–DNA interactions at the YY1, C/EBP, GATA, and GATE1-3. The signaling pathway involves gp130, JAK, STAT3, p-STAT3, PI3K, AKT, NF-κB, GPCR, SOCS3, SHP2, ERK, Grb2, Shc, Ras, and Raf. ODAM: odontogenic ameloblast-associated protein, IL-6: interleukin 6, YY1: yin yang 1, C/EBP: CCAAT/enhancer-binding protein, GATA: GATA binding protein, GATE1-3: interferon-γ activated transcriptional element 1-3, gp130: glycoprotein 130, STAT3: signal transducer and activator of transcription 3, p-STAT3: phosphorylated STAT3, PI3K: phosphatidylinositol 3-kinase, NF-κB: nuclear factor-kappa B, GPCR: G protein-coupled receptor.

Article Snippet: Twelve hours after transfection, the cells were switched to a serum-free medium for 12 hours and then treated with one of the following inhibitors for 30 minutes: 5 μM protein kinase C inhibitor H7 (Seikagaku Corporation, Tokyo, Japan), 100 nM protein kinase A inhibitor KT5720 (Sigma-Aldrich), 5 μM mitogen-activated protein kinase kinase 1/2 (MEK1/2) inhibitor U0126 (Promega), 10 μM phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002 (Calbiochem, San Diego, CA, USA), 100 nM NF-κB inhibitor triptolide (Tocris Bioscience, Bristol, UK), 1 μM glycoprotein 130 (gp130) inhibitor SC144 (MedChemExpress, Monmouth Junction, NJ, USA), 10 μM STAT1 inhibitor fludarabine (NSC 118218; Selleck Chemicals, Houston, TX, USA), or 10 μM STAT3 inhibitor stattic (Axon Medchem, Reston, VA, USA).

Techniques: Gene Expression, Binding Assay

Journal: Cell reports

Article Title: Functional genomics identifies N -acetyllactosamine extension of complex N -glycans as a mechanism to evade lysis by natural killer cells

doi: 10.1016/j.celrep.2024.114105

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Dead Cell Removal Kit , Miltenyi Biotec , 130-090-101.

Techniques: Virus, Cell Isolation, Recombinant, Plasmid Preparation, Selection, CRISPR, Glycoproteomics, Knock-Out, Software

Journal: The Journal of Neuroscience

Article Title: Involvement of Spinal IL-6 Trans -Signaling in the Induction of Hyperexcitability of Deep Dorsal Horn Neurons by Spinal Tumor Necrosis Factor-Alpha

doi: 10.1523/JNEUROSCI.4159-15.2016

Figure Lengend Snippet: Recording of neuronal activity and reduction of TNF-α-induced spinal hyperexcitability by sgp130 or anti-IL-6R. A, Experimental setup. Responses of spinal cord neurons in the deep dorsal horn with knee and ankle input were recorded in vivo. Innocuous (innoc.) and noxious (nox.) pressure were applied to the knee, ankle, and paw. Each stimulus lasted for 15 s, followed by 15 s without stimulation. Substances were applied to the surface of the spinal cord at the recording site. B, Comparison of the increases of the neuronal responses to innocuous and noxious mechanical stimulation of the knee and ankle joints for 2 h after spinal application of TNF-α (n = 11) or TNF-α plus sgp130 (n = 8), shown as mean number of action potentials per 15 s (mean APs/15 s) ± SEM for intervals of 30 min. The baseline (BL) responses before TNF-α or TNF-α plus sgp130 are set to zero. C, Comparison of the neuronal responses to innocuous and noxious knee and ankle stimulation in absolute values at baseline and 90–120 min (2 h) after TNF-α treatment and TNF-α plus sgp130 treatment. D, Comparison of the increases of the neuronal responses to innocuous and noxious mechanical stimulation of the knee and ankle joints for 2 h after spinal application of TNF-α (n = 11, same as in B) or TNF-α plus anti-IL-6R (n = 7). The gray lines display the effect of TNF-α plus the isotype control antibody (n = 3, no statistics). E, Comparison of the neuronal responses to innocuous and noxious knee and ankle stimulation in absolute values at baseline and 90–120 min (2 h) after TNF-α plus anti-IL-6R and TNF-α plus IgG2b. In B and D, asterisks with arrow indicate the start of a significant increase of responses or a significant increase of responses at the specific time points (p < 0.05 by Wilcoxon matched-pairs signed-rank test); plus signs indicate significant difference between the groups at the specific time point (p < 0.05 by Mann–Whitney U test). In C, asterisks indicate a significant increase (p < 0.05, Wilcoxon matched-pairs signed-rank test).

Article Snippet: Interestingly, endogenous sgp130 is present at high concentrations in blood and CSF ( Padberg et al., 1999 ).

Techniques: Activity Assay, In Vivo, Comparison, Control, MANN-WHITNEY

Journal:

Article Title: Porphyromonas gingivalis lipopolysaccharide induces tumor necrosis factor-α and interleukin-6 (IL-6) secretion and CCL25 gene expression in mouse primary gingival cell lines: IL-6-driven activation of CCL2

doi:

Figure Lengend Snippet: (A-C) CCL2 and TNFα levels in cell-free supernatants 24 hr after stimulation with rIL-6, rIL-6 plus sIL-6R, rIL-6 plus sIL-6R plus sgp130, sgp130, or PBS; data from 3 independent sets of experiments showing an increase in CCL2 secretion from gingival cells stimulated with rIL-6 plus sIL-6R; rIL-6 plus sIL-6R plus sgp130; or sgp130 compared to PBS control cultures. (D) Neutralization of sgp130 by anti-gp130 antibody causes an increase in CCL2 secretion. (E) A gingival cell line stained with sgp130 indicated that about one-fifth of the cells express gp130. (F) Cell supernatants used for CCL2 assays had no detectable TNFα production.

Article Snippet: Reagents used in this study included ultrapure P. gingivalis LPS (Invitrogen; San Diego; CA) and E. coli LPS (Sigma); recombinant IL-6 (rIL-6) (<0.01 ng endotoxin per μg cytokine (eBioscience; San Diego, CA); sIL-6R (<0.0 EU endotoxin per 1 μg cytokine receptor) (R&D Systems; Minneapolis, MN); two preparations of sgp130: recombinant human sgp130 (<1.0 EU endotoxin per 1 μg receptor) (R&D Systems), and recombinant mouse sgp130/Fc chimera (<1.0 ng endotoxin per 1 μg receptor) (R&D systems).

Techniques: Control, Neutralization, Staining

Journal:

Article Title: Porphyromonas gingivalis lipopolysaccharide induces tumor necrosis factor-α and interleukin-6 (IL-6) secretion and CCL25 gene expression in mouse primary gingival cell lines: IL-6-driven activation of CCL2

doi:

Figure Lengend Snippet: Stimulatory effects on CCL2 secretion from gingival cell lines

Article Snippet: Reagents used in this study included ultrapure P. gingivalis LPS (Invitrogen; San Diego; CA) and E. coli LPS (Sigma); recombinant IL-6 (rIL-6) (<0.01 ng endotoxin per μg cytokine (eBioscience; San Diego, CA); sIL-6R (<0.0 EU endotoxin per 1 μg cytokine receptor) (R&D Systems; Minneapolis, MN); two preparations of sgp130: recombinant human sgp130 (<1.0 EU endotoxin per 1 μg receptor) (R&D Systems), and recombinant mouse sgp130/Fc chimera (<1.0 ng endotoxin per 1 μg receptor) (R&D systems).

Techniques:

Journal: Nature Communications

Article Title: Interleukin-6 trans-signaling is a candidate mechanism to drive progression of human DCCs during clinical latency

doi: 10.1038/s41467-020-18701-4

Figure Lengend Snippet: a MCF 10A cells were cultured as spheres in the absence ( n = 18) or presence of IL6 ( n = 18), an IL6-blocking antibody ( n = 6) or hyper-IL6 ( n = 6). b hTERT-HME1 were cultured as spheres in the absence ( n = 3) or presence ( n = 3) of hyper-IL6. c HMECs were cultured as spheres without or with IL6, with an IL6 blocking antibody or hyper-IL6. n = 3 patients, each patient analyzed in triplicate. d hTERT-HME1-EGFR∆746-750 cells were cultured as spheres in the absence or presence of HIL6 (each n = 12). e MCF 10A cells were cultured as spheres without ( n = 6) or with IL6 ( n = 6) and IL6 plus sgp130-Fc at indicated concentrations (each n = 6). f Sphere formation of hTERT-HME1-EGFR∆746-750 in the absence ( n = 10) or presence of an anti-IL6 antibody ( n = 9) or with sgp130-Fc at indicated concentrations (each n = 12). Cumulative data of three experiments. P values in a , c , f : one-way ANOVA with Dunnett’s multiple comparisons test (post hoc); b , d two-sided Student’s t-test; e one-way ANOVA with Tukey’s multiple comparisons test (post hoc); asterisks indicate significance between groups (**** p < 0.0001). All error bars correspond to standard deviation (Mean ± SD). See Supplementary Table 1 for patient/sample-ID allocation.

Article Snippet: MCF-10A cells were cultured as mammospheres in the presence or absence of 10 ng/ml IL-6 (Sigma-Aldrich, Germany), 10 ng/ml IL-6 + 0.1 ng/ml recombinant human sgp130-Fc (R&D Systems, Germany) or 20 ng/ml HIL6 (a kind gift of S. Rose-John, Christian-Albrechts-University, Germany) for 12 and 24 h. Cells were seed in triplicates for all conditions and time points.

Techniques: Cell Culture, Blocking Assay, Standard Deviation

Journal: Pathophysiology

Article Title: Preeclampsia Status Controls Interleukin-6 and Soluble IL-6 Receptor Release from Neutrophils and Endothelial Cells: Relevance to Increased Inflammatory Responses

doi: 10.3390/pathophysiology28020013

Figure Lengend Snippet: Production of interleukin 6 (IL-6), soluble IL-6R (sIL-6R), and soluble gp130 (sgp130) by neutrophils and endothelial cells from normal and preeclamptic (PE) pregnancies. ( A ) Production of IL-6, sIL-6R, and sgp130 by neutrophils (n = 6) and endothelial cells (n = 5) from normal pregnant women. Open symbol: 2 h of culture, and solid symbol: 6 h of culture, ** p < 0.01: 6 h vs. 2 h. ( B ) Comparison of sIL-6R production by neutrophils (normal n = 6; PE n = 5), and IL-6 and sgp130 production by endothelial cells (normal n = 6; PE n = 5) from normal vs. PE pregnancies. Open symbol: normal and solid symbol: PE, * p < 0.05 and ** p < 0.01: PE vs. normal at 6 h. ( C ) Production of IL-6, sIL-6R, and sgp130 in co-cultured neutrophils and endothelial cells. Freshly isolated neutrophils (2 × 10 6 cells) were added into cultured endothelial cells (5 × 10 5 cells/per well in 6 well plate). Both neutrophils and endothelial cells were from normal pregnancy. Results were from five independent experiments. Open symbol: 2 h of culture and solid symbol: 6 h of culture, * p < 0.05 and ** p < 0.01: 6 h vs. 2 h, respectively.

Article Snippet: DuoSet ELISA development kits of IL-6 (DY206), sIL-6R (DY227), and sgp130 (DY228) were purchased from R&D systems (Minneapolis, MN, USA).

Techniques: Comparison, Cell Culture, Isolation

Journal: Pathophysiology

Article Title: Preeclampsia Status Controls Interleukin-6 and Soluble IL-6 Receptor Release from Neutrophils and Endothelial Cells: Relevance to Increased Inflammatory Responses

doi: 10.3390/pathophysiology28020013

Figure Lengend Snippet: Proposed differences in IL-6 classic signaling and IL-6 trans-signaling in neutrophils and endothelial cells. IL-6 classic signaling in neutrophils ( A ) and trans-signaling in endothelial cells ( B ) in unstimulated condition. A: IL-6 classic signaling in neutrophils. Neutrophils have both IL-6R and gp130. IL-6 binds to IL-6R and then initiates gp130 trans-signaling. B: IL-6 trans-signaling in endothelial cells. Endothelial cells have gp130, but not IL-6R. sIL-6R/IL-6 complex binds to gp130 and then initiates gp130 trans-signaling. C and D: Proposed dysregulation of IL-6 classic signaling in neutrophils ( C ) and trans-signaling in endothelial cells ( D ) in preeclamptic condition. Increased sgp130 production and increased sgp130/sIL-6R/IL-6 complex formation could prevent IL-6 to bind to IL-6R in neutrophils ( C ) and block sIL-6R/IL-6 to bind to gp130 in ECs, as a result, blocks gp130 trans-signaling.

Article Snippet: DuoSet ELISA development kits of IL-6 (DY206), sIL-6R (DY227), and sgp130 (DY228) were purchased from R&D systems (Minneapolis, MN, USA).

Techniques: Blocking Assay

Journal: bioRxiv

Article Title: Characterization of cytotrophoblast cell population dynamics throughout pregnancy

doi: 10.1101/2025.01.22.634235

Figure Lengend Snippet: BCAM is a marker for CTBs possessing progenitor capacity within the villous chorion. (A) Immunostaining of BCAM (red) and ITGA6 (green) in term placental tissue. Scale bar, 500 µm and 50 µm (zoom). (B) Immunostaining of BCAM (red) and CGB (green) in term placental tissue. Scale bar, 500 µm. (C) Brightfield images of trophoblast organoid cultures derived from BCAM + cells isolated from term placental tissue at day 7 and day 14. Scale bars, 200 µm. (D) Immunostaining of KRT7 (purple) and GDF15 (yellow) in trophoblast organoids derived from BCAM + cells at day 14. Scale bar, 200 µm.

Article Snippet: Cell suspensions were incubated with 5 μL of anti-BCAM PE-conjugated antibody (Miltenyi Biotech, 130-126-539) in the dark for 10 min at 4°C.

Techniques: Marker, Immunostaining, Derivative Assay, Isolation

Journal: bioRxiv

Article Title: Characterization of cytotrophoblast cell population dynamics throughout pregnancy

doi: 10.1101/2025.01.22.634235

Figure Lengend Snippet: Colocalization of BCAM and LAMA5 suggests contact of progenitor CTBs with basal lamina. (A) Schematic cross section of chorionic villi within the placenta. CTB; cytotrophoblast, STB; syncytiotrophoblast. (B) Electron microscopy images of term placental tissue. Blue arrowheads indicate the syncytium with multiple nuclei, green squares indicate cytotrophoblast, and red arrowheads indicate the basal lamina. Scale bars, 5 µm (top) and 1 µm (bottom). (C) STRING protein network analysis shows top 3 interactors of BCAM with interaction scores. Thickness of lines indicate confidence. (D) Immunostaining of BCAM (red) and LAMA5 (green) in term placental tissue. Scale bar, 500 µm. (E) Immunostaining of BCAM (red) in trophoblast organoids at day 14. Scale bar, 200 µm. Quantification of BCAM intensity profiles measured across multiple line scans (n = 3) per organoid (n = 10). Trend analysis is performed using orthogonal polynomial fitting (dashed line).

Article Snippet: Cell suspensions were incubated with 5 μL of anti-BCAM PE-conjugated antibody (Miltenyi Biotech, 130-126-539) in the dark for 10 min at 4°C.

Techniques: Electron Microscopy, Immunostaining

Journal: bioRxiv

Article Title: Characterization of cytotrophoblast cell population dynamics throughout pregnancy

doi: 10.1101/2025.01.22.634235

Figure Lengend Snippet: Proteomic profiling of decellularized villi across gestation. (A) Ridgeline plot showing average BCAM expression in villous CTB cells throughout gestation. (B) Bar plots showing protein abundance of BCAM and LAMA5 in decellularized villi from first trimester (n = 3) and term (n = 3) placentas. P-values are calculated using a Welch’s t-test. (C) Abundance of basal lamina-associated proteins in decellularized villi from term placentas (n = 3). (D) Venn diagram showing the number of unique or shared proteins in decellularized villi from first trimester and term placentas. Proteins present in all samples (n = 3 each) at either or both gestational times were retained. (E) Vulcano plot showing differential protein abundance of the 2043 proteins shared between decellularized villi from first and term placentas. (F) Fold change of the top 20 differentially abundant proteins in decellularized villi from either first trimester or term placentas. (G) Ridgeline plot showing average HMGA1 expression in villous CTB cells throughout gestation. (H) Bar plot showing protein abundance of HMGA1 in decellularized villi from first trimester (n = 3) and term (n = 3) placentas. P-value is calculated using a Welch’s t-test.

Article Snippet: Cell suspensions were incubated with 5 μL of anti-BCAM PE-conjugated antibody (Miltenyi Biotech, 130-126-539) in the dark for 10 min at 4°C.

Techniques: Expressing, Quantitative Proteomics