Journal: Journal of Interferon & Cytokine Research

Article Title: Identification of 2′-5′-Oligoadenylate Synthetase-Like Gene in Goose: Gene Structure, Expression Patterns, and Antiviral Activity Against Newcastle Disease Virus

doi: 10.1089/jir.2015.0167

Figure Lengend Snippet: GoOASL mRNA expression levels in PBMCs stimulated by virus pathogens (A) and agonists (B). The mRNA levels of goOASL were quantified by RT-qPCR. GAPDH was used as housekeeping gene. Data were represented as the mean ± SD (n = 3). The statistical analysis was performed in GraphPad Prism using unpaired 2-tailed t-tests: *P < 0.05; **P < 0.01; ***P < 0.001. The cell density was adjusted as 1 × 106/mL, Virus pathogens were GPV (10−6.6 EID50/0.2 mL), H9N2 AIV (7.14 × 1012.64 copies/mL), and DTMUV (6.3 × 10−6 TCID50/mL), with 50 μL to stimulate cells for 6 h, respectively. Agonists, including LPS, R848, Poly (I: C), and ODN2006, the final concentration of stimulus were 25, 5, 30, and 50 μg/mL correspondingly.

Article Snippet: Agonists are R848 (MedChem Express), LPS (Invivogen), Poly (I: C) (Sigma), and ODN2006 (Sangon).

Techniques: Expressing, Virus, Quantitative RT-PCR, Concentration Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Kinetic of RelA activation controls magnitude of TLR-mediated IL-12p40 induction.

doi: 10.4049/jimmunol.0802560

Figure Lengend Snippet: FIGURE 5. Only TLR2 and TLR4 ligands, but not ligands of other TLRs, induce serine 536 phosphorylation of RelA. A, BMDCs were stimulated with CpG-DNA (1 M) and LPS (100 ng/ml). After the indicated times, mRNA was isolated and the expression of IL-12p40 (continuous line) and IB (dotted line) was quantified by quantitative RT-PCR. The relative expression of IL-12p40 mRNA and IB in comparison to the mRNA encoding the housekeeping gene -actin is shown. Mean SD; n 3. B, BMDCs were stimulated with CpG-DNA (CpG; 1 M) and LPS (100 ng/ml) for the indicated times. Similar amounts of cellular extracts were blotted with Abs for IB and RelA phosphorylated at serine 536 (pS536RelA). C, BMDCs were stimulated with CpG-DNA (CpG; 1 M), LPS (100 ng/ml), Pam3CSK4 (P3C; 10 g/ml), R848 (1 g/ml), polyinosinic-polycytidylic acid (PIC; 50 g/ml), or imiquimod (IQ; 20 g/ml) for 10 min. Similar amounts of cellular extracts were blotted with Abs for RelA phosphorylated at serine 536 (pS536RelA) and -actin. D and E, BMDCs of MyD88 knockout (MyD) and wild-type (WT) mice (D) and LPS2 (LP2) and wild-type (WT) mice (E) were stimulated as indicated with CpG-DNA (1 M) and/or LPS (100 ng/ml) for 10 min. Similar amounts of cellular extracts (10 g/ml) were blotted with Abs for phospho-S536-RelA and -actin. Results are representative of three independent experiments.

Article Snippet: Pam3CSK4 was obtained from Alexis; R848, polyinosinic-polycytidylic acid, R-FSL, flagellin, and imiquimod were from InvivoGen; NF- B/RelA, RelB, and c-Rel Abs were obtained from Santa Cruz Biotechnology; Abs against I B , phospho-ERK, phospho-JNK, phospho-p38, phospho-I B and phospho-S536-RelA were purchased from Cell Signaling Technology.

Techniques: Phospho-proteomics, Isolation, Expressing, Quantitative RT-PCR, Comparison, Knock-Out

Journal: NPJ Vaccines

Article Title: A KIF20A-based thermosensitive hydrogel vaccine effectively potentiates immune checkpoint blockade therapy for hepatocellular carcinoma

doi: 10.1038/s41541-024-01060-2

Figure Lengend Snippet: a TEM image of K/R Lip . Scalebar: 100 nm. b Hydrated diameter of K/R Lip and ICG-K/R Lip . c Zeta potentials of K/R Lip and ICG- K/R Lip . d Hydrated diameter and Zeta potentials K/R Lip on the 1, 3, 5 and 7 days. e R848 release rate of K/R Lip at the pH of 7.4 and 5.6. f KIF20A release rate of K/R Lip at the pH of 7.4 and 5.6. g Photographs of K/R Lip @Gel at 25 °C and 37 °C. h ESEM image of K/R Lip @Gel at 37 °C. i Liposomes (K/R Lip ) detected with ESEM (Red arrows). j Rheological analysis quantifying storage modulus (G’) and loss modulus (G”) of K/R Lip @Gel at different temperatures. k Rheological analysis of gel stability for 6 min. l Absorbance spectra of ICG and ICG-K/R Lip @Gel. m Fluorescence spectra of ICG and ICG- K/R Lip @Gel. n NIR-II fluorescence images of mice injected with small molecule ICG, nanoscale ICG-K/R Lip and hydrogel ICG-K/R Lip @Gel. o Quantification of NIR-II fluorescence intensity at different time points after injections.

Article Snippet: Then, 2 mg R848 (Aladdin Scientific, China) was added into 10 mL stock solution.

Techniques: Liposomes, Fluorescence, Injection

Journal: Inflammation

Article Title: Toll-like Receptors 1, 3 and 7 Activate Distinct Genetic Features of NF-κB Signaling and γ-Protocadherin Expression in Human Cardiac Fibroblasts

doi: 10.1007/s10753-025-02238-z

Figure Lengend Snippet: Concentration of proinflammatory cytokines secreted from human cardiac fibroblasts following activation of TLR1, TLR3 or TLR7 . Cytokine concentrations were measured in conditioned medium following treatment of cardiac fibroblasts with TLR1 agonist Pam3CSK4, TLR3 agonist Poly I:C or TLR7 agonist R848 and compared to medium from untreated cells (controls). The TLR4 agonist LPS was used as a reference stimulant. The concentrations for each sample were normalized to the total protein content of that sample and expressed as pg/mg cell protein. Bars show data as the mean and standard deviation (SD). n = 7-8. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 (one way ANOVA). ( a ) IFNg = Interferon gamma, ( b ) IL-1b = Interleukin-1 beta, ( c ) IL-2 = Interleukin-2, ( d ) IL-4 = Interleukin-4, ( e ) IL-6 = Interleukin-6, ( f ) IL-8 = Interleukin-8, ( g ) IL-10 = Interleukin-10, ( h ) IL-12p70 = Interleukin-12 p70 ( i ) IL-13 = Interleukin-13, ( j ) TNFa = Tumor necrosis factor alpha

Article Snippet: Cells were treated for 24 h with or without 100 ng/ml TLR1 agonist PamCSK4 (cat# NBP2-25,297, Novus Biologicals, Centennial, CO), 100 μg/mL TLR3 agonist Poly I:C (cat# NBP2-25,288, Novus Biologicals, Centennial, CO), 5 μg/mL TLR7 agonist R848 (cat# NBP2-26,231, Novus Biologicals, Centennial, CO) or 10 ng/ml lipopolysaccharide (LPS) derived from E.coli (Invivogen, San Diego, CA).

Techniques: Concentration Assay, Activation Assay, Standard Deviation

Journal: Inflammation

Article Title: Toll-like Receptors 1, 3 and 7 Activate Distinct Genetic Features of NF-κB Signaling and γ-Protocadherin Expression in Human Cardiac Fibroblasts

doi: 10.1007/s10753-025-02238-z

Figure Lengend Snippet: γ-protocadherin gene levels in human cardiac fibroblasts following activation of TLR1, TLR3 or TLR7 . Bar graph displaying relative expression levels of γ-protocadherin genes. The dashed line indicates gene expression levels of untreated cardiac fibroblasts (controls), bars represent gene expression levels of fibroblasts treated with TLR1 agonist Pam3CSK4, TLR3 agonist Poly I:C or TLR7 agonist R848. Bars show data as the mean and standard deviation (SD). n = 8, * p -value < 0.05 (one way ANOVA)

Article Snippet: Cells were treated for 24 h with or without 100 ng/ml TLR1 agonist PamCSK4 (cat# NBP2-25,297, Novus Biologicals, Centennial, CO), 100 μg/mL TLR3 agonist Poly I:C (cat# NBP2-25,288, Novus Biologicals, Centennial, CO), 5 μg/mL TLR7 agonist R848 (cat# NBP2-26,231, Novus Biologicals, Centennial, CO) or 10 ng/ml lipopolysaccharide (LPS) derived from E.coli (Invivogen, San Diego, CA).

Techniques: Activation Assay, Expressing, Gene Expression, Standard Deviation

Journal: F&S science

Article Title: Vascular alterations impede fragile tolerance to pregnancy in type 1 diabetes

doi: 10.1016/j.xfss.2022.02.001

Figure Lengend Snippet: Cytokine analysis reveals increased interleukin-6 (IL-6) levels in NOD litters. (A) Selected cytokines from an unbiased cytokine array showing levels in amniotic fluid in picograms per milliliter. (B) Graphical representation of the levels of IL-6 family members from cytokine array (n = 3 per mouse pairing). NOD♀NOD♂ and NOD♀B6♂ litters have increases in IL-6, leukemia inhibitory factor, and monocyte chemotactic protein-1 levels. (C) The increase in the IL-6 level in amniotic fluid from NOD mice is confirmed via enzyme-linked immunosorbent assay, where each point represents amniotic fluid from 1 mouse uterus. (D) The placentas from B6♀NOD♂ and NOD♀B6♂ litters are stimulated with R848 and αCD3/αCD28 overnight and then incubated with GolgiStop for 4 hours to capture cytokine secretion. Interleukin-6 secretion from CD4+, Foxp3+, B220+, CD11b+, and CD31+ cells shows that the most IL-6 secretion is coming from CD31+ endothelial cells. (E) Percentage of CD4+, Foxp3+, B220+, CD11b+, and CD31+ of IL-6-secreting cells, where each point represents 1 placenta. Of IL-6-secreting cells, CD31+ cells had the largest percentage in both B6♀NOD♂ and NOD♀B6♂ litters. (F) The histogram of all placentas per B6♀NOD♂ litter (blues) and NOD♀B6♂ litter (pinks) shows that on average, NOD♀B6♂CD31+ cells have an increase in stored IL-6. IL = interleukin; NOD = nonobese diabetic; SSW = side scatter width; TNFα= tumor necrosis factor-α.

Article Snippet: Manually dissociated placentas were plated overnight in 5% Dulbecco’s Modified Eagle’s Medium cell culture media (with 10% FBS, penicillin/streptomycin, and β -mercaptoethanol) with 1 μ M of R848 (#73782 STEMCELL Technologies, Vancouver, Canada) and 8 μ g/mL of α CD3/ α CD28 (BD #553057 and #553294).

Techniques: Enzyme-linked Immunosorbent Assay, Incubation