human specific il 1β  (Elabscience Biotechnology)

Journal: International Journal of Molecular Sciences

Article Title: CD14 + -Monocytes Exposed to Apolipoprotein CIII Express Tissue Factor

doi: 10.3390/ijms24032223

Figure Lengend Snippet: Expression of TNFα, IL-1β and IL-6 mRNAs and protein release by human CD14 + -monocytes exposed to ApoCIII. Human CD14 + -monocytes, purified as described in , were incubated for 3 h ( A – C ) or 20 h ( D – F ) in the absence (medium) or presence of 50 µg/mL ultrapure ApoCIII or, as a control, in 100 ng/mL LPS. Total RNA was then extracted and examined for TNFα ( A ), IL-1β ( B ) and IL-6 ( C ) mRNA expression by RT-qPCR. Gene expression is depicted as mean normalized expression (MNE) units after normalization to GAPDH mRNA (mean ± SEM, n = 7–9). Monocyte-derived supernatants were collected, and TNFα ( D ), IL-1β ( E ) and IL-6 ( F ) levels were measured by ELISA. Results are expressed as the mean value ± SEM of 3 independent experiments. Statistical analysis by unpaired t-test with Welch correction refers to ApoCIII-treated versus untreated monocytes, * p < 0.05.

Article Snippet: Cytokine concentrations in cell-free supernatants were measured by commercial ELISA kits specific for human TNFα, IL-1β and IL-6 (Mabtech) according to the manufacturer’s instructions.

Techniques: Expressing, Purification, Incubation, Control, Quantitative RT-PCR, Gene Expression, Derivative Assay, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Sciences

Article Title: CD14 + -Monocytes Exposed to Apolipoprotein CIII Express Tissue Factor

doi: 10.3390/ijms24032223

Figure Lengend Snippet: Effect of sera with measured concentrations of ApoCIII on TNFα and IL-6 release by PBMCs. Panel A : PBMCs isolated from buffy coats as reported in were cultured for 3 h with the addition of ApoCIII-sera in the presence or absence of 60 µg/mL ApoCIII-neutralizing rAbs. After treatment, sera with known concentrations of ApoCIII (ApoCIII-sera) and PBMC-derived supernatants were collected and analyzed for TNFα and IL-6 content by ELISA. Data are expressed as mean± SEM (n = 8) and were analyzed by the Kruskal–Wallis test followed by Dunn’s multiple comparison test.. Panel B displays two scatter plots showing the correlation between TNFα or IL-6 release by PBMCs and the ApoCIII concentrations of the sera that were used as stimuli. The r-value determined by Pearson correlation is displayed together with the p -value.

Article Snippet: Cytokine concentrations in cell-free supernatants were measured by commercial ELISA kits specific for human TNFα, IL-1β and IL-6 (Mabtech) according to the manufacturer’s instructions.

Techniques: Isolation, Cell Culture, Derivative Assay, Enzyme-linked Immunosorbent Assay, Comparison

Journal: International Journal of Molecular Sciences

Article Title: Modulatory Effects of Fractalkine on Inflammatory Response and Iron Metabolism of Lipopolysaccharide and Lipoteichoic Acid-Activated THP-1 Macrophages

doi: 10.3390/ijms23052629

Figure Lengend Snippet: ELISArray determination of pro-inflammatory cytokine production of E. coli or P. aeruginosa LPS and S. aureus LTA and/or fractalkine-treated THP-1 cells. The arrays were carried out using pooled cell culture supernatants obtained from the same treatment types of four independent experiments ( n = 4). In the case of co-treatments, both the LPS or LTA and fractalkine were added at the same time to the cells. Supernatants were collected after 24 h of treatments. The columns show the optical density of the average Il-1β ( A ), IL-2 ( B ), IL-6 ( C ), IL-8 ( D ), TNFα ( E ) and GM-CSF ( F ) production of THP-1 cells. Abbreviations: F5—fractalkine 5 ng/mL; F10—fractalkine 10 ng/mL; LPS EC— E. coli lipopolysaccharide; LPS PA— P. aeruginosa lipopolysaccharide; LTA SA— S. aureus lipoteichoic acid.

Article Snippet: The IL-6, IL-1β and TNF-α pro-inflammatory cytokine concentrations of the culture media were determined with ELISA kits specific for human IL-1β, IL-6, and TNFα (Thermo Fisher Scientific Inc., Waltham, MA, USA).

Techniques: Cell Culture

Journal: International Journal of Molecular Sciences

Article Title: Modulatory Effects of Fractalkine on Inflammatory Response and Iron Metabolism of Lipopolysaccharide and Lipoteichoic Acid-Activated THP-1 Macrophages

doi: 10.3390/ijms23052629

Figure Lengend Snippet: ELISA measurement of IL-1β ( A ), IL-6 ( B ) and TNFα ( C ) pro-inflammatory cytokine secretion of E. coli or P. aeruginosa LPS and S. aureus LTA and/or fractalkine-treated THP-1 cells. Pro-inflammatory cytokine secretions were determined using human IL-1β, IL-6 and TNFα specific ELISA kits according to the manufacturer’s instructions. The bars represent mean values and error bars represent standard deviation (SD) for three independent determinations ( n = 3). ELISA measurements were made in triplicate in each independent experiment. Statistical significance was determined by two-way ANOVA (considering the number of the categorical variables) followed by Scheffe’s post hoc test. Asterisks indicate p < 0.05 compared to the control. Cross marks p < 0.05 compared to the fractalkine treatment. Double cross shows p < 0.05 compared to the LPS or LTA treatments, respectively. Abbreviations: F5—fractalkine 5 ng/mL; F10—fractalkine 10 ng/mL; LPS EC— E. coli lipopolysaccharide; LPS PA— P. aeruginosa lipopolysaccharide; LTA SA— S. aureus lipoteichoic acid.

Article Snippet: The IL-6, IL-1β and TNF-α pro-inflammatory cytokine concentrations of the culture media were determined with ELISA kits specific for human IL-1β, IL-6, and TNFα (Thermo Fisher Scientific Inc., Waltham, MA, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Standard Deviation, Control

Journal: International Journal of Molecular Sciences

Article Title: Modulatory Effects of Fractalkine on Inflammatory Response and Iron Metabolism of Lipopolysaccharide and Lipoteichoic Acid-Activated THP-1 Macrophages

doi: 10.3390/ijms23052629

Figure Lengend Snippet: Cell signalling pathways regulated by CX3CR1 and TLR receptors and regulation of iron metabolism of THP-1 cells. ( A ) Fractalkine binds to CX3CR1 and activates NFκB and MAPK pathways. The same signalling pathways are regulated by TLRs. NFκB transcription factors regulate the transcription of Nrf2, STAT3, HO-1 and pro-inflammatory cytokines including IL-1β, IL-6 and TNFα. Meanwhile, Nrf2, which is inhibited by Keap-1, provides feedback on NFκB and HO-1 transcription. MAPK also modifies the activity of Nrf2 and STAT3 transcription factors by phosphorylation. PSTAT3 and pro-inflammatory cytokines increase HAMP transcription. Maturation of hepcidin is regulated by A1AT. TfR1 and DMT-1 function as iron importers and Fp act as iron exporter transmembrane protein. Fp is controlled by hepcidin. FTH and FTMT work as the cytosolic and mitochondrial iron storage proteins. ( B ) Alterations of the levels of signalling proteins and iron metabolism-related proteins in the presence of soluble fractalkine in E. coli LPS activated THP-1 cells. ( C ) Alterations of the levels of signalling proteins and iron metabolism-related proteins in the presence of soluble fractalkine in P. aeruginosa LPS activated THP-1 cells. ( D ) Alterations of the levels of signalling proteins and iron metabolism related proteins in the presence of soluble fractalkine in S. aureus LTA activated THP-1 cells. Green colour marks elevation, red colour indicates reduction in the expression levels of the examined proteins.

Article Snippet: The IL-6, IL-1β and TNF-α pro-inflammatory cytokine concentrations of the culture media were determined with ELISA kits specific for human IL-1β, IL-6, and TNFα (Thermo Fisher Scientific Inc., Waltham, MA, USA).

Techniques: Activity Assay, Phospho-proteomics, Expressing

Journal: PLoS Pathogens

Article Title: Human NAIP/NLRC4 and NLRP3 inflammasomes detect Salmonella type III secretion system activities to restrict intracellular bacterial replication

doi: 10.1371/journal.ppat.1009718

Figure Lengend Snippet: WT, NAIP -/- clone #12, and two independent clones of NLRC4 -/- THP-1 monocyte-derived macrophages were primed with 100 ng/mL Pam3CSK4 for 16 hours. Cells were then treated with PBS (Mock), PA alone, LFnFlaA 310–475 alone (LFnFlaA), LFnPrgJ alone, LFnYscF alone, PA+LFnFlaA 310–475 (FlaTox), PA+LFnPrgJ (PrgJTox), or PA+LFnYscF (YscFTox) for 6 hours (A, C). As a control, cells were primed with 500 ng/mL LPS for 4 hours and treated with 10 μM nigericin for 6 hours (B, D). Release of IL-1β into the supernatant was measured by ELISA. ns–not significant, * p < 0.05, ** p < 0.01, **** p < 0.0001 by Šídák’s multiple comparisons test (A), or by unpaired t-test (B), or by Dunnett’s multiple comparisons test (C, D). Error bars represent the standard deviation of triplicate wells from one experiment. Data shown are representative of at least three independent experiments.

Article Snippet: Primary antibodies specific for human IL-1β (#8516; R&D Systems) and β-actin (4967L; Cell Signaling) and HRP-conjugated secondary antibodies anti-mouse IgG (F00011; Cell Signaling) and anti-rabbit IgG (7074S; Cell Signaling) were used.

Techniques: Clone Assay, Derivative Assay, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: PLoS Pathogens

Article Title: Human NAIP/NLRC4 and NLRP3 inflammasomes detect Salmonella type III secretion system activities to restrict intracellular bacterial replication

doi: 10.1371/journal.ppat.1009718

Figure Lengend Snippet: WT, NAIP -/- clone #12, and two independent clones of NLRC4 -/- THP-1 monocyte-derived macrophages were primed with 100 ng/mL Pam3CSK4 for 16 hours. Cells were then infected with PBS (Mock), WT S . Typhimurium, or Δ sipB S . Typhimurium at an MOI = 20 for 6 hours. Release of IL-1β (A, B) and IL-18 (C, D) into the supernatant were measured by ELISA. Immunoblot analysis was performed on supernatants (sup) for mature IL-1β and on lysates for pro–IL-1β and β-actin as a loading control. ns–not significant, *** p < 0.001, **** p < 0.0001 by Šídák’s multiple comparisons test (A, C) or Dunnett’s multiple comparisons test (B, D). Error bars represent the standard deviation of triplicate wells from one experiment. Data shown are representative of at least three independent experiments.

Article Snippet: Primary antibodies specific for human IL-1β (#8516; R&D Systems) and β-actin (4967L; Cell Signaling) and HRP-conjugated secondary antibodies anti-mouse IgG (F00011; Cell Signaling) and anti-rabbit IgG (7074S; Cell Signaling) were used.

Techniques: Clone Assay, Derivative Assay, Infection, Enzyme-linked Immunosorbent Assay, Western Blot, Standard Deviation

Journal: PLoS Pathogens

Article Title: Human NAIP/NLRC4 and NLRP3 inflammasomes detect Salmonella type III secretion system activities to restrict intracellular bacterial replication

doi: 10.1371/journal.ppat.1009718

Figure Lengend Snippet: (A, C, E) WT, NAIP -/- , or NLRC4 -/- THP-1 monocyte-derived macrophages were primed with 100 ng/mL Pam3CSK4 for 16 hours. One hour prior to infection, cells were treated with 1 μM MCC950, a chemical inhibitor of the NLRP3 inflammasome, or DMSO as a control. Cells were then infected with PBS (Mock), WT S . Typhimurium, or Δ sipB S . Typhimurium at an MOI = 20 for 6 hours. (B, D) As a control, cells were primed with 500 ng/mL LPS for 4 hours and treated with 10 μM nigericin for 6 hours. (A–D) Release of IL-1β into the supernatant was measured by ELISA. (E) Immunoblot analysis was performed on supernatants (sup) for mature IL-1β and on lysates for pro–IL-1β and β-actin as a loading control. ns–not significant, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 by Tukey’s multiple comparisons test (A, C) or by Šídák’s multiple comparisons test (B, D). Error bars represent the standard deviation of triplicate wells from one experiment. Data shown are representative of at least three independent experiments.

Article Snippet: Primary antibodies specific for human IL-1β (#8516; R&D Systems) and β-actin (4967L; Cell Signaling) and HRP-conjugated secondary antibodies anti-mouse IgG (F00011; Cell Signaling) and anti-rabbit IgG (7074S; Cell Signaling) were used.

Techniques: Derivative Assay, Infection, Enzyme-linked Immunosorbent Assay, Western Blot, Standard Deviation

Journal: PLoS Pathogens

Article Title: Human NAIP/NLRC4 and NLRP3 inflammasomes detect Salmonella type III secretion system activities to restrict intracellular bacterial replication

doi: 10.1371/journal.ppat.1009718

Figure Lengend Snippet: WT, NAIP -/- (A–C) and NLRC4 -/- (D–F) THP-1 monocyte-derived macrophages were treated with siRNA targeting a control scrambled siRNA, siRNA targeting CASP4 (A, D) or CASP5 (B, E), or siRNA targeting both CASP4 and CASP5 (C, F) for 48 hours. Cells were primed with 100 ng/mL Pam3CSK4 for 16 hours. Cells were then infected with PBS (Mock) or WT S . Typhimurium at an MOI = 20 for 6 hours. Release of IL-1β into the supernatant was measured by ELISA. As a control, cells were transfected with LPS. ns–not significant, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 by Tukey’s multiple comparisons test. Error bars represent the standard deviation of triplicate wells from one experiment. Data shown are representative of at least three independent experiments.

Article Snippet: Primary antibodies specific for human IL-1β (#8516; R&D Systems) and β-actin (4967L; Cell Signaling) and HRP-conjugated secondary antibodies anti-mouse IgG (F00011; Cell Signaling) and anti-rabbit IgG (7074S; Cell Signaling) were used.

Techniques: Derivative Assay, Infection, Enzyme-linked Immunosorbent Assay, Transfection, Standard Deviation

Journal: PLoS Pathogens

Article Title: Human NAIP/NLRC4 and NLRP3 inflammasomes detect Salmonella type III secretion system activities to restrict intracellular bacterial replication

doi: 10.1371/journal.ppat.1009718

Figure Lengend Snippet: WT, NAIP -/- (A, B), and NLRC4 -/- (C, D) THP-1 monocyte-derived macrophages were primed with 100 ng/mL Pam3CSK4 for 16 hours. One hour prior to infection, cells were treated with 1 μM MCC950, a chemical inhibitor of the NLRP3 inflammasome, or DMSO as a control. Cells were then infected with PBS (Mock) or WT S . Typhimurium at an MOI = 20. Cells were lysed at the indicated time points and bacteria were plated to calculate CFU. (A, C) CFU/well of bacteria at 6 hpi (B, D) Fold change in CFU/well of bacteria at indicated time point, relative to 2 hpi CFU/well. ns–not significant, *** p < 0.001, **** p < 0.0001 by Dunnett’s multiple comparisons test (A, C) or Tukey’s multiple comparisons test (B, D). Error bars represent the standard deviation of triplicate wells from one experiment. Data shown are representative of at least three independent experiments.

Article Snippet: Primary antibodies specific for human IL-1β (#8516; R&D Systems) and β-actin (4967L; Cell Signaling) and HRP-conjugated secondary antibodies anti-mouse IgG (F00011; Cell Signaling) and anti-rabbit IgG (7074S; Cell Signaling) were used.

Techniques: Derivative Assay, Infection, Bacteria, Standard Deviation

Journal: PLoS Pathogens

Article Title: Human NAIP/NLRC4 and NLRP3 inflammasomes detect Salmonella type III secretion system activities to restrict intracellular bacterial replication

doi: 10.1371/journal.ppat.1009718

Figure Lengend Snippet: WT and NAIP -/- THP-1 monocyte-derived macrophages were seeded on glass coverslips and primed with 100 ng/mL Pam3CSK4 for 16 hours. One hour prior to infection, cells were treated with 1 μM MCC950, a chemical inhibitor of the NLRP3 inflammasome, or DMSO as a control. Cells were then infected with PBS (Mock) or WT S . Typhimurium expressing GFP at an MOI = 20. Cells were fixed at the indicated time points and stained for DAPI to label DNA (blue). The proportion of infected cells containing GFP-expressing Stm (green) and the number of bacteria per cell were scored by fluorescence microscopy. (A) Representative images from 6hpi are shown. Scale bar represents 10 μm. (B, C) Each small dot represents one infected cell. 150 infected cells were scored for each condition (50 infected cells per coverslip). Bars represent the mean from each condition. (B) Number of bacteria/cell at 2 hpi. (C) Number of bacteria/cell at 6 hpi. ns–not significant, **** p < 0.0001 by Tukey’s multiple comparisons test (B). Data shown are representative of at least three independent experiments.

Article Snippet: Primary antibodies specific for human IL-1β (#8516; R&D Systems) and β-actin (4967L; Cell Signaling) and HRP-conjugated secondary antibodies anti-mouse IgG (F00011; Cell Signaling) and anti-rabbit IgG (7074S; Cell Signaling) were used.

Techniques: Derivative Assay, Infection, Expressing, Staining, Bacteria, Fluorescence, Microscopy

Journal: PLoS Pathogens

Article Title: Human NAIP/NLRC4 and NLRP3 inflammasomes detect Salmonella type III secretion system activities to restrict intracellular bacterial replication

doi: 10.1371/journal.ppat.1009718

Figure Lengend Snippet: (A) Primary hMDMs from four healthy human donors were infected with PBS (Mock), WT Listeria ( WT Lm ) , Listeria expressing PrgJ (Lm + PrgJ), SsaI (Lm + SsaI), or SsaG (Lm + SsaG) for 16 hours at MOI = 5. Release of IL-18 into the supernatant was measured by ELISA. Each dot represents the mean of individual donors derived from triplicate wells. The grey bar represents the mean of all donors. (B) WT, NAIP -/- , and NLRC4 -/- THP-1 monocyte-derived macrophages were primed with 100 ng/mL Pam3CSK4 for 16 hours. Cells were treated with PBS (Mock), WT Listeria ( WT Lm ) , Listeria expressing PrgI (Lm + PrgI), or Listeria expressing SsaG (Lm + SsaG) for 6 hours at MOI = 20. Release of IL-1β into the supernatant was measured by ELISA. (C) WT, NAIP -/- , and NLRC4 -/- THP-1 monocyte-derived macrophages were primed with 100 ng/mL Pam3CSK4 for 16 hours. Cells were then treated with PBS (Mock), PA alone, LFnSsaG, or PA+LFnSsaG (SsaGTox) for 6 hours. Release of IL-1β into the supernatant was measured by ELISA. ns–not significant, *** p < 0.001, **** p < 0.0001 paired t-test (A) or by Dunnett’s multiple comparisons test (B, C). Data shown are representative of at least three independent experiments.

Article Snippet: Primary antibodies specific for human IL-1β (#8516; R&D Systems) and β-actin (4967L; Cell Signaling) and HRP-conjugated secondary antibodies anti-mouse IgG (F00011; Cell Signaling) and anti-rabbit IgG (7074S; Cell Signaling) were used.

Techniques: Infection, Expressing, Enzyme-linked Immunosorbent Assay, Derivative Assay

Journal: PLoS Pathogens

Article Title: Human NAIP/NLRC4 and NLRP3 inflammasomes detect Salmonella type III secretion system activities to restrict intracellular bacterial replication

doi: 10.1371/journal.ppat.1009718

Figure Lengend Snippet: WT, NAIP -/- , and NLRC4 -/- THP-1 monocyte-derived macrophages were primed with 100 ng/mL Pam3CSK4 for 16 hours. Cells were then infected with PBS (Mock) or Δ prgIfliCfljB S . Typhimurium at an MOI = 20. (A, B) Cells were lysed at the indicated time points and bacteria were plated to calculate CFU. Fold change in CFU/well of bacteria at indicated time point, relative to 2 hpi CFU/well. (C, D) Release of IL-1β was measured at 24hpi by ELISA. ns–not significant, **** p < 0.0001 by Šídák’s multiple comparisons test.

Article Snippet: Primary antibodies specific for human IL-1β (#8516; R&D Systems) and β-actin (4967L; Cell Signaling) and HRP-conjugated secondary antibodies anti-mouse IgG (F00011; Cell Signaling) and anti-rabbit IgG (7074S; Cell Signaling) were used.

Techniques: Derivative Assay, Infection, Bacteria, Enzyme-linked Immunosorbent Assay

Journal: BMC Complementary Medicine and Therapies

Article Title: Anti-inflammatory effect of lavender ( Lavandula angustifolia Mill.) essential oil prepared during different plant phenophases on THP-1 macrophages

doi: 10.1186/s12906-021-03461-5

Figure Lengend Snippet: Real-time PCR gene primers used in the experiments

Article Snippet: The concentrations of IL-6, IL-1β, IL-8 and TNFα were determined in triplicate in each independent experiment using human IL-6, IL-1β, IL-8 and TNFα specific ELISA kits (Thermo Fisher Scientific Inc., Waltham, MA) according to the instructions of the manufacturer.

Techniques: Real-time Polymerase Chain Reaction, Sequencing

Journal: BMC Complementary Medicine and Therapies

Article Title: Anti-inflammatory effect of lavender ( Lavandula angustifolia Mill.) essential oil prepared during different plant phenophases on THP-1 macrophages

doi: 10.1186/s12906-021-03461-5

Figure Lengend Snippet: mRNA expression ( A,C,E,G ) and protein ( B,D,F,H ) levels of pro-inflammatory cytokines IL-6, IL-8, IL-1β and TNFα after linalool and LEOs treatments of THP-1 cells. THP-1 cells were treated with 500-fold diluted linalool and LEOs prepared at the beginning and at the end of flowering period. DMSO treated cells were used as a control of the essential oil treated cells. Real Time PCR was performed with SYBR green protocol using pro-inflammatory cytokine specific primers. β-actin was used as housekeeping gene for the normalization. Relative expression of controls was considered as 1. Pro-inflammatory cytokine secretions were determined using IL-6, IL-8, IL-1β and TNFα specific ELISA kits according to the manufacturer’s instructions. The bars represent mean values and error bars represent standard deviation (SD) for three independent determinations ( n = 3). Real Time PCR and ELISA measurements were carried out in triplicate in each independent experiment. Asterisks indicate p < 0.05 compared to the control. The diagrams were created using the same logical structure and the different treatments appeared in the same sequence in each figure in order to improve traceability of the results

Article Snippet: The concentrations of IL-6, IL-1β, IL-8 and TNFα were determined in triplicate in each independent experiment using human IL-6, IL-1β, IL-8 and TNFα specific ELISA kits (Thermo Fisher Scientific Inc., Waltham, MA) according to the instructions of the manufacturer.

Techniques: Expressing, Real-time Polymerase Chain Reaction, SYBR Green Assay, Enzyme-linked Immunosorbent Assay, Standard Deviation, Sequencing

Journal: BMC Complementary Medicine and Therapies

Article Title: Anti-inflammatory effect of lavender ( Lavandula angustifolia Mill.) essential oil prepared during different plant phenophases on THP-1 macrophages

doi: 10.1186/s12906-021-03461-5

Figure Lengend Snippet: Effects of linalool, LEOs prepared at the beginning and at the end of the flowering period and NFκB inhibitors on mRNA ( A,C,E,G ) and protein ( B,D,F,H ) levels of pro-inflammatory cytokines IL-6, IL-8, IL-1β and TNFα after P. aeruginosa LPS pretreatment. THP-1 cells were pretreated with 100 ng/mL LPS for 24 h then 500-fold diluted linalool and LEOs or 5 μM luteolin or ACHP were added to the pretreated THP-1 cells for an additional 24 h. DMSO treated cells were used as control of the treated cells. Real Time PCR was carried out with SYBR green protocol using pro-inflammatory cytokine specific primers. β-actin was used as housekeeping gene for the normalization and relative expression of controls was regarded as 1. Pro-inflammatory cytokine secretions were determined using specific ELISA kits according to the manufacturer’s protocols. The columns represent mean values and error bars represent standard deviation (SD) for three independent determinations ( n = 3). Real Time PCR and ELISA measurements were carried out in triplicate in each independent experiment. Asterisks indicate p < 0.05 compared to the control. Crosses show p < 0.05 compared to the LPS treatment. Double cross represent statistical significance ( p < 0.05) compared to luteolin treatment. Number sign marks p < 0.05 compared to ACHP treatment. The diagrams were created using the same logical structure and the different treatments appeared in the same sequence in each figure in order to improve traceability of the results

Article Snippet: The concentrations of IL-6, IL-1β, IL-8 and TNFα were determined in triplicate in each independent experiment using human IL-6, IL-1β, IL-8 and TNFα specific ELISA kits (Thermo Fisher Scientific Inc., Waltham, MA) according to the instructions of the manufacturer.

Techniques: Real-time Polymerase Chain Reaction, SYBR Green Assay, Expressing, Enzyme-linked Immunosorbent Assay, Standard Deviation, Sequencing

Journal: BMC Complementary Medicine and Therapies

Article Title: Anti-inflammatory effect of lavender ( Lavandula angustifolia Mill.) essential oil prepared during different plant phenophases on THP-1 macrophages

doi: 10.1186/s12906-021-03461-5

Figure Lengend Snippet: Effects of linalool and LEOs and NFκB inhibitors pretreatments on mRNA ( A,C,E,G ) and protein ( B,D,F,H ) levels of pro-inflammatory cytokines IL-6, IL-8, IL-1β and TNFα. THP-1 cells were pretreated with 500-fold diluted linalool, LEOs and 5 μM luteolin or ACHP for 24 h then 100 ng/mL P. aeruginosa LPS was added to the pretreated cells for 24 h. DMSO administration was used as a control of the treated cells. Real Time PCR was performed with SYBR green protocol using specific primers. β-actin was used as housekeeping gene for the normalization and the relative expression of controls was considered as 1. Pro-inflammatory cytokine secretions were determined using IL-6, IL-8, IL-1β and TNFα specific ELISA kits according to the manufacturer’s instructions. The bars represent mean values and error bars represent standard deviation (SD) for three independent determinations ( n = 3). Real Time PCR and ELISA measurements were carried out in triplicate in each independent experiment. Asterisks indicate p < 0.05 compared to the control. Crosses show p < 0.05 compared to the LPS treatment. Double cross represents p < 0.05 compared to luteolin treatment. Number sign marks p < 0.05 compared to ACHP treatment. The diagrams were created using the same logical structure and the different treatments appeared in the same sequence in each figure in order to improve traceability of the results

Article Snippet: The concentrations of IL-6, IL-1β, IL-8 and TNFα were determined in triplicate in each independent experiment using human IL-6, IL-1β, IL-8 and TNFα specific ELISA kits (Thermo Fisher Scientific Inc., Waltham, MA) according to the instructions of the manufacturer.

Techniques: Real-time Polymerase Chain Reaction, SYBR Green Assay, Expressing, Enzyme-linked Immunosorbent Assay, Standard Deviation, Sequencing