Journal: Scientific Reports

Article Title: A potential peptide derived from cytokine receptors can bind proinflammatory cytokines as a therapeutic strategy for anti-inflammation

doi: 10.1038/s41598-018-36492-z

Figure Lengend Snippet: Surface property analysis of preferable binding poses for KCF18 to proinflammatory cytokines. KCF18 is represented as an orange-colored and green-colored loop structure. Residues around the binding interface are labeled and are shown as sticks. Surface charge distribution of KCF18 binding to cytokines was calculated using the Poisson–Boltzmann equation. Blue color corresponds to positive and red color to negative electrostatic potential. ( A ) KCF18–TNF-α, ( C ) KCF18–IL-1β, and ( E ) KCF18–IL-6 complexes. Surface lipophilicity distribution of KCF18 binding to cytokines, in which blue color represents hydrophilic residues, whereas green color represents hydrophobic residues. ( B ) KCF18–TNF-α, ( D ) KCF18–IL-1β, and ( F ) KCF18–IL-6 complexes.

Article Snippet: The recombinant proteins of TNF-α (catalog # 1130-01), IL-1β (catalog # 1110-01B), and IL-6 (catalog # 1110-06) were purchased from the GOLDBIO.COM, USA.

Techniques: Binding Assay, Labeling

Journal: Scientific Reports

Article Title: A potential peptide derived from cytokine receptors can bind proinflammatory cytokines as a therapeutic strategy for anti-inflammation

doi: 10.1038/s41598-018-36492-z

Figure Lengend Snippet: SPR analysis for KCF18 binding to proinflammatory cytokines. ( A ) KCF18 was injected over TNF-α immobilized on the CM5 sensor chip. As the concentration of KCF18 increased, the measured response for the binding of KCF18 to TNF-α also increased, indicating that binding was concentration dependent. ( B ) For the steady–state interaction, a binding isotherm was generated to determine the equilibrium K D and R max for the binding of KCF18 to cytokine TNF-α, which were found to be 60.9 μM and 197.1 RU, respectively. ( C ) KCF18 was injected over IL-6 immobilized on the CM5 sensor chip. The measured response for the binding of KCF18 to IL-6 also increased with the concentration of KCF18, similar to the profile of the binding of KCF18 to TNF-α. ( D ) The kinetic analysis of binding isotherm was also performed to determine the equilibrium dissociation K D and R max for KCF18 binding to cytokine IL-6, which were found to be 111.5 μM and 210.3 RU, respectively.

Article Snippet: The recombinant proteins of TNF-α (catalog # 1130-01), IL-1β (catalog # 1110-01B), and IL-6 (catalog # 1110-06) were purchased from the GOLDBIO.COM, USA.

Techniques: Binding Assay, Injection, Concentration Assay, Generated

Journal: Scientific Reports

Article Title: A potential peptide derived from cytokine receptors can bind proinflammatory cytokines as a therapeutic strategy for anti-inflammation

doi: 10.1038/s41598-018-36492-z

Figure Lengend Snippet: KCF18 inhibits various cytokine-induced monocyte adhesion to HMEC-1. HMEC-1 cells were pretreated with various concentrations of KCF18 or ( D ) peptide CF25 for 1 hour and were then stimulated with 20 ng/mL ( A ) TNF-α, (B) IL-1β, or ( C ) IL-6 for 18 hours. Adhesion of fluorescent THP-1 cells was photographed by fluorescent microscopy, and fluorescence intensity was calculated. “Control” cells were only incubated with the culture medium (without peptides). Values are mean ± SD from three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001 as compared with control; # P < 0.05, ## P < 0.01, ### P < 0.001 and #### P < 0.0001 as compared with cells stimulated with cytokines in the absence of peptides.

Article Snippet: The recombinant proteins of TNF-α (catalog # 1130-01), IL-1β (catalog # 1110-01B), and IL-6 (catalog # 1110-06) were purchased from the GOLDBIO.COM, USA.

Techniques: Microscopy, Fluorescence, Incubation

Journal: Scientific Reports

Article Title: A potential peptide derived from cytokine receptors can bind proinflammatory cytokines as a therapeutic strategy for anti-inflammation

doi: 10.1038/s41598-018-36492-z

Figure Lengend Snippet: KCF18 inhibits cytokine-induced transmigration of monocytes. HMEC-1 cells were pretreated with various concentrations of KCF18 for 1 hour and then stimulated with 20 ng/mL ( A ) TNF-α, ( B ) IL-1β, or ( C ) IL-6 for 18 hours; thereafter, THP-1 cells were allowed to transmigrate through the HMEC-1 monolayer. Values are mean ± SD from three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001 as compared with control; # P < 0.05, ## P < 0.01, ### P < 0.001 and #### P < 0.0001 as compared with cells stimulated with cytokines in the absence of KCF18.

Article Snippet: The recombinant proteins of TNF-α (catalog # 1130-01), IL-1β (catalog # 1110-01B), and IL-6 (catalog # 1110-06) were purchased from the GOLDBIO.COM, USA.

Techniques: Transmigration Assay

Journal: Scientific Reports

Article Title: A potential peptide derived from cytokine receptors can bind proinflammatory cytokines as a therapeutic strategy for anti-inflammation

doi: 10.1038/s41598-018-36492-z

Figure Lengend Snippet: Peptides KCF18 and CF25 affect the expression of cytokine-induced TNF-α in HMEC-1 and THP-1 cells. TNF-α mRNA levels induced by TNF-α ( A and D ), IL-1β ( B and E ), or IL-6 ( C and F ) in HMEC-1 ( A–C ) and THP-1 ( D–F ) were determined using qPCR assays, as described in Materials and Methods. GAPDH cDNA was used as an internal control. Values are mean ± SD of mRNA levels relative to those for GAPDH from three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001 as compared with control; # P < 0.05, ## P < 0.01, ### P < 0.001 and #### P < 0.0001 as compared with cells stimulated with cytokines in the absence of the peptides.

Article Snippet: The recombinant proteins of TNF-α (catalog # 1130-01), IL-1β (catalog # 1110-01B), and IL-6 (catalog # 1110-06) were purchased from the GOLDBIO.COM, USA.

Techniques: Expressing

Journal: Scientific Reports

Article Title: A potential peptide derived from cytokine receptors can bind proinflammatory cytokines as a therapeutic strategy for anti-inflammation

doi: 10.1038/s41598-018-36492-z

Figure Lengend Snippet: Superposition of peptide KCF18-cytokine system and receptor-cytokine system. ( A ) KCF18-TNF-α and TNF-α-TNFR1 ( B ) KCF18-IL-1β and IL-1β-IL1R ( C ) KCF18-IL-6 and IL-6-IL6R. Gray color is cytokine, cyan color is receptor, purple color is the region selected from receptor to compose the peptide KCF18, and peptide KCF18 is composed of three color short peptide (orange: TNFR1; red: IL1R; green: IL6R).

Article Snippet: The recombinant proteins of TNF-α (catalog # 1130-01), IL-1β (catalog # 1110-01B), and IL-6 (catalog # 1110-06) were purchased from the GOLDBIO.COM, USA.

Techniques:

Journal: The Journal of Biological Chemistry

Article Title: Interferon-γ induces the cell surface exposure of phosphatidylserine by activating the protein MLKL in the absence of caspase-8 activity

doi: 10.1074/jbc.RA118.007161

Figure Lengend Snippet: The MLKL protein forms a trimer in PS-exposing MEFs treated with IFN-γ. A and B, primary WT MEFs (A) and iC8KO-RIPK3 MEFs (B) were treated with 10 ng/ml IFN-γ and 50 μm z-VAD-fmk (IZ) and with 10 ng/ml IFN-γ, respectively, for the indicated periods. Cell lysates were resolved on reducing SDS-PAGE and analyzed by Western blotting with anti-phosphorylated MLKL (pMLKL), anti-MLKL, and anti-actin antibodies. C and D, cell lysates of primary WT MEFs treated with IZ (C) or with 10 ng/ml TNFα, 1 μg/ml cycloheximide, and 50 μm z-VAD-fmk (TCZ) (D) for the indicated periods were resolved on reducing or nonreducing SDS-PAGE and analyzed by Western blotting with anti-MLKL and anti-actin antibodies. E and F, iC8KO-RIPK3 MEFs (E) or primary WT MEFs, iC8KO-RIPK3 MEFs, and iC8KO-RIPK3 MEFs expressing shMlkl#2 (F) were treated with IFN-γ for the indicated periods. Cell lysates were resolved on reducing or nonreducing SDS-PAGE and analyzed by Western blotting with anti-MLKL and anti-actin antibodies. *, nonspecific bands. Representative Western blotting data of more than three experiments are shown.

Article Snippet: z-VAD-fmk (Peptide Institute), mouse and human IFN-γ (Pepro Tech), mouse TNFα (ProSpec), Staurosporine (Alomone Labs), Necrostatin-1 (Santa Cruz Biotechnology), anti-mouse TNFα antibody (506310, Biolegend), Smac mimetics (A11928, AdooQ Bioscience), and NSA (Toronto Research Chemicals) were used as a pan-caspase inhibitor, necroptosis inducer, necroptosis inducer, apoptosis inducer, RIPK1 inhibitor, TNFα inhibitor, IAP inhibitor, and human MLKL inhibitor, respectively.

Techniques: SDS Page, Western Blot, Expressing

Journal: Medicine

Article Title: Determination of diagnostic and predictive parameters for vertical mandibular invasion in patients with lower gingival squamous cell carcinoma: A retrospective study

doi: 10.1097/MD.0000000000032206

Figure Lengend Snippet: Immunohistochemical staining.

Article Snippet: IL-6 , Rockland Inc. , 1:2000 , Rabbit polyclonal antibody , • Inflammatory cytokines released by cancer cells. • Since stimulation by IL-6 and TNF-α induces bone-resorbing multinucleated giant cells, it is possible that some mechanisms of the bone-resorptive effects of IL-6 are not mediated by the RANK/RANKL pathway..

Techniques: Immunohistochemical staining, Staining, Binding Assay, Activity Assay, Expressing

Journal: Frontiers in Cardiovascular Medicine

Article Title: CTRP3 attenuates myocardial lipotoxicity via suppression of lipid accumulation, inflammation, apoptosis, and mitochondrial oxidative stress

doi: 10.3389/fcvm.2025.1575929

Figure Lengend Snippet: (A,B) Compared to the high-fat diet (HFD) group, supplementation with CTRP3 significantly improves cardiac function in mice, as evidenced by enhanced left ventricular ejection fraction. (C,D) Histological examination of myocardial tissue reveals substantial lipid droplet accumulation in the HFD group. However, CTRP3 supplementation reduces lipid droplet accumulation, improves myocardial tissue structure, and alleviates the degree of fibrosis. (E–M) Immunofluorescence results indicate that CTRP3 supplementation mitigates myocardial inflammation, apoptosis, and oxidative stress in mice. (N–Q) Serum ELISA assays demonstrate that levels of triglycerides (TG), total cholesterol (TCHO), inflammatory markers, apoptotic factors, and oxidative stress indicators are significantly improved in the CTRP3 group compared to the HFD group ( n ≥ 3, p < 0.05).

Article Snippet: The recombinant CTRP3 protein (rCTRP3, Catalog Number: CSB-EP883621HU) was commercially procured from Wuhan Cusabio Biotech Co., Ltd. (Hubei, China).

Techniques: Immunofluorescence, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Cardiovascular Medicine

Article Title: CTRP3 attenuates myocardial lipotoxicity via suppression of lipid accumulation, inflammation, apoptosis, and mitochondrial oxidative stress

doi: 10.3389/fcvm.2025.1575929

Figure Lengend Snippet: (A–F) Lipid testing results demonstrate that palmitic acid (PA) induces lipid accumulation in myocardial cells, while CTRP3 supplementation significantly reduces lipid droplet accumulation. (G) CTRP3 intervention improves the expression of genes related to fatty acid uptake, fatty acid oxidation, and lipid efflux in myocardial cells ( n ≥ 3, p < 0.05). (H) Quantitative PCR (q-PCR) results indicate that CTRP3 mitigates inflammation, apoptosis, and oxidative stress in myocardial cells stimulated by PA ( n ≥ 3, p < 0.05).

Article Snippet: The recombinant CTRP3 protein (rCTRP3, Catalog Number: CSB-EP883621HU) was commercially procured from Wuhan Cusabio Biotech Co., Ltd. (Hubei, China).

Techniques: Expressing, Real-time Polymerase Chain Reaction

Journal: Frontiers in Cardiovascular Medicine

Article Title: CTRP3 attenuates myocardial lipotoxicity via suppression of lipid accumulation, inflammation, apoptosis, and mitochondrial oxidative stress

doi: 10.3389/fcvm.2025.1575929

Figure Lengend Snippet: (A–E) RNA sequencing analysis of myocardial tissue from the control group, HFD group, and CTRP3 group indicates that CTRP3 likely exerts its effects within myocardial mitochondria, playing a role in the metabolic regulation of myocardial lipotoxicity.

Article Snippet: The recombinant CTRP3 protein (rCTRP3, Catalog Number: CSB-EP883621HU) was commercially procured from Wuhan Cusabio Biotech Co., Ltd. (Hubei, China).

Techniques: RNA Sequencing, Control

Journal: Frontiers in Cardiovascular Medicine

Article Title: CTRP3 attenuates myocardial lipotoxicity via suppression of lipid accumulation, inflammation, apoptosis, and mitochondrial oxidative stress

doi: 10.3389/fcvm.2025.1575929

Figure Lengend Snippet: (A,B) JC-1 staining results demonstrate a decrease in mitochondrial membrane potential in myocardial cells of the PA group. (C,D) Mito-Tracker staining results show a reduction in mitochondrial number in the PA group. (E) PA intervention in myocardial cells leads to decreased ATP production, while CTRP3 intervention improves ATP generation. (F) Transmission electron microscopy reveals increased mitochondrial damage and autophagy in myocardial cells following PA intervention. CTRP3 can mitigate mitochondrial damage and autophagy in myocardial cells.

Article Snippet: The recombinant CTRP3 protein (rCTRP3, Catalog Number: CSB-EP883621HU) was commercially procured from Wuhan Cusabio Biotech Co., Ltd. (Hubei, China).

Techniques: Staining, Membrane, Transmission Assay, Electron Microscopy

Journal: Oncotarget

Article Title: A novel anti-GD2/4-1BB chimeric antigen receptor triggers neuroblastoma cell killing

doi:

Figure Lengend Snippet: A. GD2 CAR T cells were capable of substantially abrogate tumor growth either versus SH-SY5Y treated with parental GFP only T cells or SH-SY5Y alone (* p = 0.036, ** p = 0.027 by t -test, mean ± SEM). Arrows represent T cell injection time points. T cells were injected locally into the tumor. B. Representative tumors for each group (first row); representative images of anti-GFP staining (second row with insets) of tumor sections obtained from mice injected with SH-SY5Y cells alone (left), SH-SY5Y+GFP only T (middle) or SH-SY5Y+GD2 CAR T cells (right); in the fourth row (with insets) TUNEL assay detecting apoptosis. Scale bars: 50 μm and 10 μm for insets. C. Fold of increase in apoptosis of GD2 CAR T cell treated specimen versus either SH-SY5Y alone or SH-SY5Y treated with GFP only T cells.

Article Snippet: Interferon gamma (IFNγ) and tumor necrosis factor related apoptosis inducing ligand (TRAIL) levels were measured in supernatants from 48-hour coculture assays, using either Human IFNγ ELISA Kit (Boster immunoleader, Fremont, CA) and Human TRAIL kit assay (R&D System Inc, Minneapolis, MN).

Techniques: Injection, Staining, TUNEL Assay

Journal: International Journal of Oncology

Article Title: Tunicamycin enhances the suppressive effects of cisplatin on lung cancer growth through PTX3 glycosylation via AKT/NF-κB signaling pathway

doi: 10.3892/ijo.2018.4650

Figure Lengend Snippet: dePTX3 suppresses the growth and migration of human lung cancer cells. (A) CCK-8 assay was used to analyze the viability of A549 and SPCA1 cells. Cells were treated with or without rhPTX3 (100 ng/ml) or dePTX3 (rhPTX3; 100 ng/ml + PNGase F; 500 U/ml) for 0-72 h. Data were presented as a percentage of viable cells. (B) Lung cancer cells treated with TM (0.01, 0.05, 0.5 and 1 µ g/ml) for 48 h. (C) Immunofluorescent staining of PTX3 (red color) and N-glycan levels (green color) in A549 and SPCA1 cells analyzed following treatment with TM (0.5 µ g/ml) for 48 h. (D) SDS-PAGE gel (12%) stained with CBB and lectinblot against PHA to analyze N-glycans in lung cancer cells following treatment with TM for 48 h and PNGase F (500 U/ml) for 2 h of incubation. (E) Western blot analysis of dePTX3 in A549 and SPCA1 cells following treatment with TM or rhPTX3 + PNGase F. The lanes are labeled as follows: lane 1, glycosylated PTX3; lane 2, deglycosylated PTX3 by TM; lane 3, deglycosylated PTX3 by PNGase F. (F) Representative images of wound healing of A549 and SPCA1 cells analyzed after treatment with rhPTX3, dePTX3 and TM (0.5 µ g/ml), followed by measurement of relative wound width (48 h average wound width divided by 0 h wound width). (G) Microscopic images and graphs of transwell migration assay of A549 and SPCA1 cells following treatment with rhPTX3, dePTX3 and TM. Bar graph values represent the means ± SEM of 3 independent experiments. * P<0.05 and ** P<0.01.

Article Snippet: Cell Counting kit-8 (CCK-8) was obtained from Beyotime Biotechnology (Shanghai, China); recombinant human PTX3 (rhPTX3) (#PROTP26022; Boster Biological Technology Co., Ltd., Fremont, CA); NF-κB inhibitor (IKK-16), PI3K inhibitor (GDC0941), MEK1/2 inhibitor, (MEK162) were purchased from Selleck Chemicals (Shanghai, China).

Techniques: Migration, CCK-8 Assay, Staining, Glycoproteomics, SDS Page, Incubation, Western Blot, Labeling, Transwell Migration Assay

Journal: International Journal of Oncology

Article Title: Tunicamycin enhances the suppressive effects of cisplatin on lung cancer growth through PTX3 glycosylation via AKT/NF-κB signaling pathway

doi: 10.3892/ijo.2018.4650

Figure Lengend Snippet: PTX3 expression in human lung cancer samples and cell lines. (A) Immunohistochemical staining of PTX3 in paired human lung cancer tissue (P1, P2, P3) and normal lung tissue (N1, N2, N3) of the same patient (magnification, ×10). (B) PTX3 level in serum from lung cancer patients and normal healthy individuals by ELISA; * P<0.05. (C) PTX3 level in the supernatant collected from A549, SPCA1 and H1299 cells was examined by ELISA. (D) Immunofluorescent staining of PTX3 protein (red color) in A549, SPCA1 and H1299 lung cancer cells. DAPI (blue color) was used to stain the nuclei. (E) mRNA expression level of PTX3 detected by qPCR in A549, SPCA1 and H1299 cells. (F) PTX3 expression level in A549, SPCA1 and H1299 cells detected by western blot analysis. GAPDH was used as an internal control.

Article Snippet: Cell Counting kit-8 (CCK-8) was obtained from Beyotime Biotechnology (Shanghai, China); recombinant human PTX3 (rhPTX3) (#PROTP26022; Boster Biological Technology Co., Ltd., Fremont, CA); NF-κB inhibitor (IKK-16), PI3K inhibitor (GDC0941), MEK1/2 inhibitor, (MEK162) were purchased from Selleck Chemicals (Shanghai, China).

Techniques: Expressing, Immunohistochemical staining, Staining, Enzyme-linked Immunosorbent Assay, Western Blot, Control

Journal: International Journal of Oncology

Article Title: Tunicamycin enhances the suppressive effects of cisplatin on lung cancer growth through PTX3 glycosylation via AKT/NF-κB signaling pathway

doi: 10.3892/ijo.2018.4650

Figure Lengend Snippet: PTX3 deglycosylation enhances the sensitivity of lung cancer cells to Cisplatin treatment. (A) suPTX3 level was detected by ELISA in the supernatant of A549 and SPCA1 cells following treatment with Cis (20 µ M) for 48 h. (B) Immunofluorescent staining of PTX3 (red color) in A549 and SPCA1 cells following treatment with Cis for 48 h. DAPI (blue color) was used to stain the nuclei. (C) Bar graph of A549 and SPCA1 cell viability analyzed following treatment with Cis and deglycosylated PTX3 (dePTX3). Each bar represents the average of 3 independent experiments. Data are expressed as a percentage of viable cells. (D) Western blot analysis of PCNA and AKT phosphorylation following treatment with Cis or dePTX3, and combined treatment for 48 h. (E) Western blot analysis for PTX3 expression in the cell lysate from control, mock and mutated PTX3 (mPTX3) in A549 and SPCA1 cells. (F) Western blot analysis of Cis, mPTX3 or Cis combined with mPTX3 and combined treatment for 48 h. The level of PCNA expression and AKT phosphorylation were examined by western blot analysis. GAPDH was used as an internal control. * P<0.05 and ** P<0.01.

Article Snippet: Cell Counting kit-8 (CCK-8) was obtained from Beyotime Biotechnology (Shanghai, China); recombinant human PTX3 (rhPTX3) (#PROTP26022; Boster Biological Technology Co., Ltd., Fremont, CA); NF-κB inhibitor (IKK-16), PI3K inhibitor (GDC0941), MEK1/2 inhibitor, (MEK162) were purchased from Selleck Chemicals (Shanghai, China).

Techniques: Enzyme-linked Immunosorbent Assay, Staining, Western Blot, Phospho-proteomics, Expressing, Control

Journal: International Journal of Oncology

Article Title: Tunicamycin enhances the suppressive effects of cisplatin on lung cancer growth through PTX3 glycosylation via AKT/NF-κB signaling pathway

doi: 10.3892/ijo.2018.4650

Figure Lengend Snippet: TM increases the sensitivity of lung cancer cells to cisplatin-induced apoptosis through deactivating AKT/NF-κB signaling pathway. (A) CCK-8 assay used to analyze the viability of A549 and SPCA1 cells treated with GDC0941 (4 µ M), IKK-16 (10 µ M), MEK162 (20 µ M) or rhPTX3 (100 ng/ml) for 48 h. (B) Western blot analysis of Bcl2 expression in A549 and SPCA1 cells, incubated with rhPTX3 (100 ng/ml) with or without NF-κB inhibitor IKK-16 (10 µ M). (C) A549 and SPCA1 cells were treated with TM (0.5 µ g/ml) + Cis (20 µ M) together or in combination. Western blot analysis of AKT, IKK, p65 phosphorylation in cytoplasmic and nuclear extracts of A549 and SPCA1 cells treated with TM, Cis or in combination. PARP was used as an internal control. (D) A549 and SPCA1 cells were double-stained with Annexin V-FITC/PI and analyzed by flow cytometry after 24 h of treatment with TM (0.5 µ g/ml), Cis (20 µ M) or combined treatment. The histogram showed the average percentage of total apoptotic cells in both A549 and SPCA1 cells. (E) Bcl2, Bax and cleaved PARP expression detected by western blot analysis following treatment with TM, Cis or in combination for 48 h. GAPDH was used as an internal control. Data represented the mean values ± SEM. * P<0.05 and ** P<0.01.

Article Snippet: Cell Counting kit-8 (CCK-8) was obtained from Beyotime Biotechnology (Shanghai, China); recombinant human PTX3 (rhPTX3) (#PROTP26022; Boster Biological Technology Co., Ltd., Fremont, CA); NF-κB inhibitor (IKK-16), PI3K inhibitor (GDC0941), MEK1/2 inhibitor, (MEK162) were purchased from Selleck Chemicals (Shanghai, China).

Techniques: CCK-8 Assay, Western Blot, Expressing, Incubation, Phospho-proteomics, Control, Staining, Flow Cytometry