Journal: Cancers

Article Title: Cancer-Associated Fibroblasts Promote Tumor Aggressiveness in Head and Neck Cancer through Chemokine Ligand 11 and C-C Motif Chemokine Receptor 3 Signaling Circuit

doi: 10.3390/cancers14133141

Figure Lengend Snippet: Characterization of CAFs and NFs obtained from clinical surgical tissues from patients with head and neck cancer (HNC) ( a ) Morphological comparisons between CAFs and NFs from a representative HNC case showed that CAFs (right panel) consisted of more cytoplasmic protrusions than NFs (left panel). Photographs were captured at 40× magnification. ( b ) Quantitative PCR (left panel) of the culture medium showed a significantly higher expression of vimentin and α-SMA in CAFs than in NFs. Western blot analysis (right panel) also demonstrated that levels of vimentin and α-SMA were significantly higher in CAFs than in NFs. ( c ) Flow cytometric analysis of the cell surface markers, CD10 and GPR77, showed a marked increase in their expression in CAFs compared to NFs. ( d ) A heat map of the gene microarray of NFs and CAFs showed that there were several differences in the expression profile of the secreted genes. The arrow indicates a marked discrepancy of the relative mRNA levels of CCL11 in CAF compared with NF. ( e ) The RT-PCR (left panel) and ELISA (right panel) analysis showed an increased expression of CCL11 in CAFs compared with that in NFs. ( f ) Western blot analysis showed that the protein level of CCL11 was higher in CAFs than in NFs in cell lysates. ( g ) Western blot analysis showed a higher CCL11 expression in CAF-CM compared to NF-CM. The asterisk indicated a significant difference (*: p < 0.05; **: p < 0.01) between experimental and control groups.

Article Snippet: CAFs and NFs supernatant CCL11concentrations were measured using the CCL11 (human) ELISA kit (R&D system) following the manufacturer’s protocol.

Techniques: Real-time Polymerase Chain Reaction, Expressing, Western Blot, Microarray, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Control

Journal: Cancers

Article Title: Cancer-Associated Fibroblasts Promote Tumor Aggressiveness in Head and Neck Cancer through Chemokine Ligand 11 and C-C Motif Chemokine Receptor 3 Signaling Circuit

doi: 10.3390/cancers14133141

Figure Lengend Snippet: CCL11 produced by CAFs causes increased migration and invasion, and the EMT of HNC cells. ( a ) Comparative analysis of the migration and invasion of HNC cells associated with CCL11. Four test groups were classified for comparative analysis of migration and invasive abilities. FaDu and NPC204 cells cultured with medium containing CAF-induced CCL11 presented greater abilities of migration and invasion, with a statistically significant difference, than three other groups: NF, NF with CCL11, and CAFs treated with CCL11 antibody. The asterisk indicates a significant difference (*: p < 0.05; **: p < 0.01) ( b ) Comparative photographs of the infiltrating behavior of FaDu and NPC204 cells in an organotypic culture in four groups seeded onto a mixture layer containing NFs or CAFs with CCL11 or CCL11 antibody. The arrow(s) indicate infiltration buds from the HNC cells seeded above. ( c ) Representative blots of the EMT-associated markers in FaDu and NPC204 cells, as observed upon Western blotting analysis in five groups, showed that treatment with CAF-conditioned medium or the application of rCCL11 decreased the expression of epithelial-type markers (E-cadherin), and increased the expression of mesenchymal-type markers (fibronectin) and EMT regulators (Snail and Twist). In addition, increased expression of invasion-related MMP2 and MMP9 was also seen in those two groups, compared with other groups. The asterisk indicates a significant difference ( p < 0.05) between experimental and control groups. Results are expressed as mean ± SD.

Article Snippet: CAFs and NFs supernatant CCL11concentrations were measured using the CCL11 (human) ELISA kit (R&D system) following the manufacturer’s protocol.

Techniques: Produced, Migration, Cell Culture, Western Blot, Expressing, Control

Journal: Cancers

Article Title: Cancer-Associated Fibroblasts Promote Tumor Aggressiveness in Head and Neck Cancer through Chemokine Ligand 11 and C-C Motif Chemokine Receptor 3 Signaling Circuit

doi: 10.3390/cancers14133141

Figure Lengend Snippet: Comparative analysis of induction of CSC properties and drug resistance in HNC cells associated with CCL11. ( a ) Four groups were classified for comparative analysis of the ability of sphere formation. Increased ability of sphere formation in two test groups of HNC cells exposed to a CAF medium and the group with treatment of rCCL11 was noted. ( b ) Flow cytometric analysis showed a significant increase in CD44 and CD44/CD24, as well as in CD133 in HNC cells exposed to rCCL11, compared to control HNC cells ( p < 0.05). ( c ) Flow cytometric analysis showed a marked increase in ALDH-1 activity in HNC cells exposed to rCCL11 compared to control HNC cells. ( d ) Western blot analysis showed that CSC-representative markers, Oct-4, Nanog, and Sox-2, were also overexpressed in addition to the increased expression of two important drug resistance genes, ABCG-2 and MDR-1 , in HNC cells exposed to rCCL11. ( e ) Treatment with Cisplatin at 24 h showed a significant increase in chemoresistance in both FaDu and NPC204 cells exposed to rCCL11 compared with control HNC cells. The asterisk indicates a significant difference (*: p < 0.05; **: p < 0.01) between experimental and control groups.

Article Snippet: CAFs and NFs supernatant CCL11concentrations were measured using the CCL11 (human) ELISA kit (R&D system) following the manufacturer’s protocol.

Techniques: Control, Activity Assay, Western Blot, Expressing

Journal: Cancers

Article Title: Cancer-Associated Fibroblasts Promote Tumor Aggressiveness in Head and Neck Cancer through Chemokine Ligand 11 and C-C Motif Chemokine Receptor 3 Signaling Circuit

doi: 10.3390/cancers14133141

Figure Lengend Snippet: CCL11 and CCR3 expression with associated signal pathway in HNC cell lines and their correlation to clinical outcomes in 104 HNC patients. ( a ) Confocal microscopic images showed CCL11 (green) localized to both cell and nuclear membranes, while CCR3 (red) localized only to the cell membrane in FaDu cells; CCL11 and CCR3 co-localized at the cell membrane (yellow). In NPC204 cells, CCL11 (green) and CCR3 (red) were found to co-localize at protrusions polarized to the cells (yellow). ( b ) Using the crisp technique, higher expression of CCR3, MMP2, and MMP3 was found in over-expressed CCL11 cloned-FaDu and NPC204 cells. Cloned CCL11-overexpressed cells were abolished by adding eotaxin siRNA or CCR3 antibody, which reversed the expression of CCR3 and invasion-related MMP2 and MMP9. ( c ) Higher phosphorylation levels of p38 MAPK and ERK were found in cloned CCL11-overexpressed FaDu and NPC204 cells and were reversed by treatment of the p38 MAPK inhibitor (SB203580) and ERK inhibitor (FR180204), respectively. The phosphorylation level of JNK was kept in low condition before and after treatment of the JNK inhibitor (SP600125). ( d ) Photomicrographs of immunohistochemical staining from tissue microarray showing CCL11 and CCR3 expression in three different representative groups of HNC patients (magnification, ×200). ( e ) Kaplan–Meier survival analysis of patients showed that overexpression of CCL11 and CCR3 were statistically associated with poor overall survival).

Article Snippet: CAFs and NFs supernatant CCL11concentrations were measured using the CCL11 (human) ELISA kit (R&D system) following the manufacturer’s protocol.

Techniques: Expressing, Membrane, Clone Assay, Phospho-proteomics, Immunohistochemical staining, Staining, Microarray, Over Expression

Journal: Cancers

Article Title: Cancer-Associated Fibroblasts Promote Tumor Aggressiveness in Head and Neck Cancer through Chemokine Ligand 11 and C-C Motif Chemokine Receptor 3 Signaling Circuit

doi: 10.3390/cancers14133141

Figure Lengend Snippet: The diagrammatic illustration demonstrates the major mechanism that CAFs secreting CCL11 promotes HNC cell migration and invasion and induces properties of drug resistance and stemness, shown as follows. CAFs in TME secret CCL11 binding to the CCR3 receptors on HNC cells via the paracrine effect. The signal induces overexpression of transcriptional factors, such as Snail and Twist, which regulate EMT and are also responsible for self-induction of CCL11 in an autocrine fashion. As a result, CCL11, via paracrine or autocrine signaling when targeting CCR3 receptors, play a functional role in the induction of EMT and CSC properties, for further tumor progression.

Article Snippet: CAFs and NFs supernatant CCL11concentrations were measured using the CCL11 (human) ELISA kit (R&D system) following the manufacturer’s protocol.

Techniques: Migration, Binding Assay, Over Expression, Functional Assay

Journal: The Journal of Experimental Medicine

Article Title: ICOS signaling limits regulatory T cell accumulation and function in visceral adipose tissue

doi: 10.1084/jem.20201142

Figure Lengend Snippet: Increased VAT-T R accumulation in the absence of ICOS signaling is associated with elevated CCR3 expression. (A) ST2 expression in splenic T R s ( n = 3–5 per group from eight independent experiments). (B) ST2 expression in CD45.1 + and CD45.2 + donor splenic T R s in WT:YF and WT:KO chimeric mice. Lines connect CD45.1 + and CD45.2 + donor splenic T R s within the same chimera ( n = 3–5 chimeric mice per group from three independent experiments). (C) Expression of CCR3 in VAT-T R s in mice ≤8 wk and >8 wk of age ( n = 3–5 per group from seven independent experiments). (D) Expression of CCR2 and CCR3 by VAT-T R s ( n = 3–5 per group from two independent experiments). DP, CCR2 + CCR3 + ; DN, CCR2 − CCR3 − . (E) Expression of indicated CCR3 ligands in total VAT normalized to Tbp as measured by qPCR (n.d. indicates not detected; n = 5 per group). (F) Schematic of in vitro culture experiments examining the impact of ICOS signaling on CCR3 expression (left). Graphs indicating fold change in T R frequency of CD4 + cells and %CCR3 + of T R s between individual culture samples stimulated (stim) with or without αICOS for 2 d (middle). Representative flow cytometry plots with frequency of CCR3 + T R s after 2 d in specified culture conditions (right; n = 1–3 per group from three independent experiments). (G) Left: Representative flow cytometry plots indicating VAT-T R frequency with or without CCL11/24 blockade. Graphs summarize T R frequencies in indicated tissues after 2 wk ( n = 3–4 per group). Mice were age matched within independent experiments and collectively; pooled data are from experiments using male mice aged 8–16 wk unless otherwise indicated. Statistical significance was determined using one-way ANOVA with Tukey’s post-test (A and C); two-tailed, paired Student’s t test for expression in donor cells within the same chimeric mouse (B); and two-tailed Student’s t test (F and G). All data are presented as mean values ± SD.

Article Snippet: Mice aged 8–10 wk were given 0.75 µg/g body weight αCCL11 and αCCL24 (MAB420 and MAB528; R&D Systems) or an equivalent amount of rat IgG (Sigma) diluted in PBS by i.p. injection on days 0, 5, and 10 and sacrificed for analysis on day 13.

Techniques: Expressing, In Vitro, Flow Cytometry, Two Tailed Test

Journal: The Journal of Experimental Medicine

Article Title: ICOS signaling limits regulatory T cell accumulation and function in visceral adipose tissue

doi: 10.1084/jem.20201142

Figure Lengend Snippet: Increased accumulation of CCR3 + T R s in the absence of ICOS signaling is specific to VAT (goes with ). (A) CCR2 expression by splenic (top) and VAT-T R s (bottom) as measured by flow cytometry ( n = 3–5 per group from two independent experiments). (B) Expression of CCR3 in splenic T R s in mice ≤8 wk and >8 wk of age ( n = 3–5 per group from seven independent experiments). (C) Expression of CCR3 by gated CD45.1 + and CD45.2 + donor VAT-T R s in chimeric mice. Graphs summarize CCR3 expression by donor T R s in VAT and spleen. Line connects point representing CD45.1 + and CD45.2 + cells within the same chimeric mouse ( n = 2–4 per group from two independent experiments). (D) CCR3 expression by tissue-localized skin T R s as measured by flow cytometry ( n = 2–4 per group from two independent experiments). (E) CCR3 expression by CD45.1 + and CD45.2 + donor T R s within the same chimeric mouse in indicated tissues. Line connects CD45.1 + and CD45.2 + cells within the same chimeric mouse ( n = 2–4 per group from two independent experiments). (F) Frequency of tissue-restricted VAT eosinophils with and without in vivo CCL11/24 blockade as measured by flow cytometry ( n = 3 or 4 mice per group). Mice were age matched within individual experiments, and pooled data are from experiments using 8–16-wk-old male mice. Statistical significance was determined using one-way ANOVA with Tukey’s post-test (A, B, and D); two-tailed, paired Student’s t test for expression in donor cells within the same chimeric mouse (C and E); and two-tailed Student’s t test (F). All data are presented as mean values ± SD. LPL, lamina propria lymphocyte.

Article Snippet: Mice aged 8–10 wk were given 0.75 µg/g body weight αCCL11 and αCCL24 (MAB420 and MAB528; R&D Systems) or an equivalent amount of rat IgG (Sigma) diluted in PBS by i.p. injection on days 0, 5, and 10 and sacrificed for analysis on day 13.

Techniques: Expressing, Flow Cytometry, In Vivo, Two Tailed Test

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Acupuncture Alleviates Menstrual Pain in Rat Model via Suppressing Eotaxin/CCR3 Axis to Weak EOS-MC Activation

doi: 10.1155/2022/4571981

Figure Lengend Snippet: The primers used for RT-qPCR analysis.

Article Snippet: The specific CCR3 agonist CCL11 (420-ME-100/CF, R&D Systems, MN, USA), which was reconstituted at 100 μ g/mL in sterile PBS (Vehicle2), was injected intraperitoneally at a dose of 10 μ g/per animal 30 min before the acupuncture.

Techniques:

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Acupuncture Alleviates Menstrual Pain in Rat Model via Suppressing Eotaxin/CCR3 Axis to Weak EOS-MC Activation

doi: 10.1155/2022/4571981

Figure Lengend Snippet: TN reduced eotaxin and histamine (HIS) levels and decreased the expression of CC chemokine receptor 3 (CCR3) and histamine H1 receptor (H1R) in CCD rats. The levels of eotaxin (a, b), HIS (c, d), and interleukin-6 (IL-6) (e, f) in uterus and plasma were detected by ELISA. The mRNA expression of CCR3 (g) and H1R (h) in the uterus was analyzed by RT-qPCR. (i–k) Representative blots (i), and protein expression levels of CCR3 (j) and H1R (k) in uterus. All data are expressed as the means ± SEM ( n = 6 rats/group). (b–f, j, k) One-way ANOVA followed by the Bonferroni post hoc test was used. (a, g, h) The Tamhane T2 post hoc test was used. ∗∗ P < 0.01 vs. Blank group; ∗ P < 0.05 vs. Blank group; △△ P < 0.01 vs. Model group; △ P < 0.05 vs. Model group.

Article Snippet: The specific CCR3 agonist CCL11 (420-ME-100/CF, R&D Systems, MN, USA), which was reconstituted at 100 μ g/mL in sterile PBS (Vehicle2), was injected intraperitoneally at a dose of 10 μ g/per animal 30 min before the acupuncture.

Techniques: Expressing, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Acupuncture Alleviates Menstrual Pain in Rat Model via Suppressing Eotaxin/CCR3 Axis to Weak EOS-MC Activation

doi: 10.1155/2022/4571981

Figure Lengend Snippet: CCR3 agonist blocked the effect of TN against CCD-induced menstrual pain. (a) The representative uterine contraction curves of all groups. The representative images of the rat uterine activities. (i–vi) Model, Model + SB328437, Model + Vehicle1, Model + CCL11 + TN, Model + Vehicle2 + TN, and Model + Vehicle1 + TN. (b) The number of contraction wave. (c) The peak-to-peak value. (d) The degree of contraction. Data were analyzed by the Tamhane T2 post hoc test and expressed as means ± SEM ( n = 6 rats/group). ∗ P < 0.05 vs. Model group; △△ P < 0.01 vs. Model + Vehicle2 + TN; △ P < 0.05 vs. Model + Vehicle2 + TN; ## P < 0.01 vs. Model + Vehicle1 + TN; # P < 0.05 vs. Model + Vehicle1 + TN.

Article Snippet: The specific CCR3 agonist CCL11 (420-ME-100/CF, R&D Systems, MN, USA), which was reconstituted at 100 μ g/mL in sterile PBS (Vehicle2), was injected intraperitoneally at a dose of 10 μ g/per animal 30 min before the acupuncture.

Techniques:

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Acupuncture Alleviates Menstrual Pain in Rat Model via Suppressing Eotaxin/CCR3 Axis to Weak EOS-MC Activation

doi: 10.1155/2022/4571981

Figure Lengend Snippet: CCR3 agonist abolished the effect of TN against CCD-induced increased ECP expression and HIS level. (a) The representative photomicrographs of ECP immunohistochemical staining of all groups in the rat uterus. (i–vi) Model, Model + SB328437, Model + Vehicle1, Model + CCL11 + TN, Model + Vehicle2 + TN, and Model + Vehicle1 + TN. (b) Histogram of IOD value of ECP in the uterus of CCD rats in each group. (c) The level of HIS in uterus was detected by ELISA. The regions are photographs at 400x magnification. The arrows indicate a positive expression of ECP. All data are expressed as the means ± SEM ( n = 6 rats/group). (b) The Tamhane T2 post hoc test was used. (c) One-way ANOVA followed by the Bonferroni post hoc test was used. ∗∗ P < 0.01 vs. Model group; △△ P < 0.01 vs. Model + Viecle2 + TN; ## P < 0.01 vs. Model + Viecle1 + TN; # P < 0.05 vs. Model + Viecle1 + TN.

Article Snippet: The specific CCR3 agonist CCL11 (420-ME-100/CF, R&D Systems, MN, USA), which was reconstituted at 100 μ g/mL in sterile PBS (Vehicle2), was injected intraperitoneally at a dose of 10 μ g/per animal 30 min before the acupuncture.

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

Journal: British Journal of Pharmacology

Article Title: Induction of mast cell accumulation by chymase via an enzymatic activity‐ and intercellular adhesion molecule‐1‐dependent mechanism

doi: 10.1111/bph.14117

Figure Lengend Snippet: The release of 5‐HT and eotaxin induced by chymase in the peritoneum of mice. (A) Various concentrations of chymase in the presence or absence of chymostatin (Chymos, 5 μg), elastase (μg) or CI A23187 (μM) were injected i.p. 10 min and 3 h before the peritoneal lavage fluids were collected for 5‐HT measurement. (B) Various concentrations of chymase in the presence or absence of chymostatin (Chymos, 5 μg), 5‐HT (μg) in the presence or absence of WAY100635 (μg) or elastase (μg) were injected i.p. 10 min and 3 h before the peritoneal lavage fluids were collected for eotaxin measurement. Chymase (μg) and CI (μM) were injected i.p. in wild‐type C57BL/6J mice and mast cell‐deficient (KitW‐4Bao) mice before peritoneal levels of 5‐HT (C) or eotaxin (D) were determined. NS was employed as a vehicle. Data are displayed as a boxplot, which indicates the median, interquartile range, the largest and smallest values. Each data group represents the results from six to seven animals. *P < 0.05 compared with the corresponding NS group. † P < 0.05 compared with the corresponding stimulus alone group.

Article Snippet: Rat monoclonal antibodies including anti‐mouse CD11a lymphocyte function‐associated antigen 1 α chain, anti‐mouse CD18 (integrin β2 chain), anti‐mouse CD62L (L‐selectin), rat IgG2a isotype standard; hamster anti‐mouse CD54 (ICAM‐1) antibody and hamster IgG1 isotype standard, recombinant mouse neutrophil elastase and mouse eotaxin and RANTES elisa kits were purchased from R&D Systems (Minneapolis, USA).

Techniques: Injection

Journal: British Journal of Pharmacology

Article Title: Induction of mast cell accumulation by chymase via an enzymatic activity‐ and intercellular adhesion molecule‐1‐dependent mechanism

doi: 10.1111/bph.14117

Figure Lengend Snippet: Mast cell accumulation in the peritoneum of mice induced by 5‐HT and chemokines. (A) 5‐HT (μg) in the presence or absence of WAY100635 (μg) was injected i.p. for 10 min or 3 h. (B) Various concentrations of eotaxin (ng) or (C) RANTES (ng) were injected i.p. for 3, 6 and 16 h. NS was employed as a vehicle. Data are displayed as a boxplot, which indicates the median, interquartile range, the largest and smallest values. Each group of data represents results from six to seven animals. *P < 0.05 compared with the corresponding NS group. † P < 0.05 compared with the corresponding stimulus alone group.

Article Snippet: Rat monoclonal antibodies including anti‐mouse CD11a lymphocyte function‐associated antigen 1 α chain, anti‐mouse CD18 (integrin β2 chain), anti‐mouse CD62L (L‐selectin), rat IgG2a isotype standard; hamster anti‐mouse CD54 (ICAM‐1) antibody and hamster IgG1 isotype standard, recombinant mouse neutrophil elastase and mouse eotaxin and RANTES elisa kits were purchased from R&D Systems (Minneapolis, USA).

Techniques: Injection

Journal: British Journal of Pharmacology

Article Title: Induction of mast cell accumulation by chymase via an enzymatic activity‐ and intercellular adhesion molecule‐1‐dependent mechanism

doi: 10.1111/bph.14117

Figure Lengend Snippet: Inhibition of mast cell accumulation induced by antibodies against cell adhesion molecules and drugs. (A) Mice were pretreated with monoclonal antibody against L‐selectin (anti‐CD62L), CD11a (anti‐CD11a), CD18 (anti‐CD18) and ICAM‐1 (anti‐CD54), respectively (all at a dose of 1 mg·kg−1), for 30 min before i.p. injection of eotaxin (5 ng) or RANTES (5 ng) for 6 h. (B) Mice were pretreated with sodium cromoglycate (Cromogl, 20 mg·kg−1) and terfenadine (Terf, 2 mg·kg−1) for 30 min before i.p. injection of eotaxin (5 ng) or RANTES (5 ng) for 6 h. NS was employed as a vehicle. Data are displayed as a boxplot, which indicates the median, interquartile range, the largest and smallest values. Each data group represents results from six to seven animals. *P < 0.05 compared with the corresponding NS group. † P < 0.05 compared with the corresponding stimulus alone group.

Article Snippet: Rat monoclonal antibodies including anti‐mouse CD11a lymphocyte function‐associated antigen 1 α chain, anti‐mouse CD18 (integrin β2 chain), anti‐mouse CD62L (L‐selectin), rat IgG2a isotype standard; hamster anti‐mouse CD54 (ICAM‐1) antibody and hamster IgG1 isotype standard, recombinant mouse neutrophil elastase and mouse eotaxin and RANTES elisa kits were purchased from R&D Systems (Minneapolis, USA).

Techniques: Inhibition, Injection

Journal: Molecular Medicine Reports

Article Title: Psoralen inhibits the inflammatory response and mucus production in allergic rhinitis by inhibiting the activator protein 1 pathway and the downstream expression of cystatin-SN

doi: 10.3892/mmr.2021.12291

Figure Lengend Snippet: PSO inhibits inflammation in IL-13-induced JME/CF15 cells. (A) A Cell Counting Kit-8 assay was performed to detect cell viability (n=5). The expression of GM-CSF was detected via (B) ELISA and (C) RT-qPCR (n=5). The expression of Eotaxin was detected via (D) ELISA and (E) RT-qPCR. (F) RT-qPCR was performed to determine the expression of IL-6 and IL-8 (n=5). (G) Western blotting was conducted to detect the expression of IL-6 and IL-8 (n=5). (H) The expression of reactive oxygen species was detected using a 2′,7′-dichlorodihydrofluorescein diacetate fluorescent prob (n=3). (I) Statistical analysis of fluorescence intensity. n=3. **P<0.01, ***P<0.001 vs. control group; # P<0.05, ## P<0.01, ### P<0.001 vs. IL-13 group. PSO, psoralen; GM-CSF, granulocyte-macrophage colony-stimulating factor; RT-qPCR, reverse transcription-quantitative PCR.

Article Snippet: Quantification of granulocyte-macrophage colony-stimulating factor (GM-CSF; cat. no. SGM00; R&D Systems, Inc.) and Eotaxin (cat. no. MME00; R&D Systems, Inc.) in cell supernatants (300 × g; 4°C; 10 min) was performed using an ELISA kit according to the manufacturer's instructions ( ).

Techniques: Cell Counting, Expressing, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Western Blot, Fluorescence, Control, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Molecular Medicine Reports

Article Title: Psoralen inhibits the inflammatory response and mucus production in allergic rhinitis by inhibiting the activator protein 1 pathway and the downstream expression of cystatin-SN

doi: 10.3892/mmr.2021.12291

Figure Lengend Snippet: PSO inhibits inflammation in IL-13-induced JME/CF15 cells by inhibiting the activator protein 1 signaling pathway. The expression of GM-CSF was detected via (A) ELISA and (B) RT-qPCR (n=5). The expression of Eotaxin was detected via (C) ELISA and (D) RT-qPCR (n=5). (E) RT-qPCR was performed to determine the expression of IL-6 and IL-8 (n=5). (F) Western blotting was performed to determine the expression of IL-6 and IL-8 (n=3). (G and H) The expression of reactive oxygen species was detected using a 2′,7′-dichlorodihydrofluorescein diacetate probe (n=3). ***P<0.001 vs. control group; ### P<0.001 vs. IL-13 group; ΔP<0.05, ΔΔP<0.01, ΔΔΔP<0.001 vs. PSO + IL-13 group; @ P<0.05, @@ P<0.01, @@@ P<0.001 vs. PMA + PSO + IL-13 group. PSO, psoralen; GM-CSF, granulocyte-macrophage colony-stimulating factor; RT-qPCR, reverse transcription-quantitative PCR.

Article Snippet: Quantification of granulocyte-macrophage colony-stimulating factor (GM-CSF; cat. no. SGM00; R&D Systems, Inc.) and Eotaxin (cat. no. MME00; R&D Systems, Inc.) in cell supernatants (300 × g; 4°C; 10 min) was performed using an ELISA kit according to the manufacturer's instructions ( ).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Western Blot, Control, Reverse Transcription, Real-time Polymerase Chain Reaction