recombinant murine il 17a  (R&D Systems)

Journal: CNS Neuroscience & Therapeutics

Article Title: IL ‐ 17A Promotes NETs Formation via the PKCζ – ERK – ROS – PAD4 Pathway in a Mouse Model of Ischemic Stroke

doi: 10.1002/cns.70825

Figure Lengend Snippet: PAD4 expression and CitH3‐positive cell ratio decreased in the peri‐infarction area of IL‐17A − / − mice. (A) Immunofluorescence staining images of the peri‐infarction area of mice. Anti‐PAD4 (green) and DAPI (blue) were used for immunofluorescence staining (scale bar = 100 μm). Green fluorescent secondary antibody was used to stain PAD4. (B) Statistical graph of PAD4 fluorescence intensity. ** p < 0.01 versus sham group, ## p < 0.01, n = 6 per group. (C) Immunofluorescence staining images of the peri‐infarction area of mice. Anti‐citH3 (green) and DAPI (blue) were used for immunofluorescence staining (scale bar = 20 μm). (D) Statistical graph of CitH3‐positive cell number. Data are expressed as mean ± standard error. Statistical results were analyzed by One‐way ANOVA. Bonfferoni test was used for comparison between groups. Images are representative of independent experimental batches. *** p < 0.001 versus sham group, ## p < 0.01, n = 6 per group.

Article Snippet: After isolation, neutrophils were cultured in RPMI‐1640 medium supplemented with 10% FBS and stimulated with recombinant murine IL‐17A (rmIL‐17A, 250 ng/mL, #CX14, Novoprotein, Suzhou, CN) for 1 h, 2 h, 4 h, and 6 h, respectively.

Techniques: Expressing, Immunofluorescence, Staining, Fluorescence, Comparison

Journal: CNS Neuroscience & Therapeutics

Article Title: IL ‐ 17A Promotes NETs Formation via the PKCζ – ERK – ROS – PAD4 Pathway in a Mouse Model of Ischemic Stroke

doi: 10.1002/cns.70825

Figure Lengend Snippet: IL‐17A regulates the expression of PAD4 through PKCζ‐ERK1/2‐ROS. (A) Flow cytometry images for the sorting and identification of neutrophils using CD11b and Ly‐6G antibodies. (B) Cellular immunofluorescence of Ly‐6G showed the purity of neutrophils. (C) Primary neutrophils were taken and Western blot was used to detect the expression of target proteins under different group stimulation conditions. rm‐IL‐17A, recombinant mouse IL‐17A; ζ‐Stat, inhibitor of PKC‐ζ; Ravoxertinib, ERK kinase inhibitor; Apocynin, NADPH oxidase inhibitor; GSK484, PAD4 inhibitor. (D) Statistical graph of PCK‐ζ. (E) Statistical graph of ERK phosphorylation level. (F) Statistical graph of PAD4 expression. (G and H) The levels of ROS were detected by DCFH‐DA probe in primary neutrophils cells under different group stimulation. The statistical graph (G) and the representative flow cytometry graph (H) were presented. Statistical results were analyzed by One‐way ANOVA. Bonferroni test was used for comparison between groups, *** p < 0.001, ### p < 0.001, ## p < 0.01, # p < 0.05, n = 6 per group.

Article Snippet: After isolation, neutrophils were cultured in RPMI‐1640 medium supplemented with 10% FBS and stimulated with recombinant murine IL‐17A (rmIL‐17A, 250 ng/mL, #CX14, Novoprotein, Suzhou, CN) for 1 h, 2 h, 4 h, and 6 h, respectively.

Techniques: Expressing, Flow Cytometry, Immunofluorescence, Western Blot, Recombinant, Phospho-proteomics, Comparison

Journal: bioRxiv

Article Title: Mycobacterium tuberculosis suppresses protective Th17 responses during infection through multiple mechanisms

doi: 10.1101/2025.05.08.652811

Figure Lengend Snippet: (A-C) WT B6 mice were aerosol infected with WT M. tuberculosis Erdman strain and evaluated at 21 days post infection (dpi) for (A) bacterial burdens in lungs by CFU from lungs (B) IFN-γ + CD4 + T cells and (C) IL-17a + CD4 + T cells from lungs using flow cytometry. Cytokines were measured using ICS after restimulation with Ag85b or ESAT-6 peptide pools. (D-E) Mice were infected and analyzed at 170 dpi for (D) IFN-γ + CD4 + T cells and (E) IL-17a + CD4 + T cells in lungs using flow cytometry. (F-H) WT B6 mice were either aerosol infected or intranasally infected with WT M. tuberculosis Erdman strain and evaluated 28 dpi for (F) bacterial burdens in lungs by CFU, (G) IFN-γ + CD4 + T cells and (H) IL-17a + CD4 + T cells in lungs by flow cytometry. Cytokines were measured using ICS after restimulation with ESAT-6 peptide pool. (I) WT B6 mice were aerosol infected with WT M. tuberculosis and treated with PBS control, recombinant murine IL-17a (IL-17a) or recombinant murine IL-17a fused to murine serum albumin (MSA-IL-17a) daily either using the intranasal (i.n.) or intraperitoneal (i.p.) routes between days 11 to 20 post infection. CFU in lungs was measured at 21 dpi. Results are representative of at least 2 biological replicates. For all graphs, each dot represents an individual mouse. *p<0.05, **p<0.01 (unpaired nonparametric Mann-Whitney U test).

Article Snippet: Mice were intranasally administered 1 μg endotoxin-free recombinant murine IL-17a (R&D systems, 7956-ML-100/CF) in 40 μL sterile PBS daily or 40 μL of vehicle control daily, or intraperitoneally administered 2 μg endotoxin-free recombinant murine IL-17a in 100 μL sterile PBS daily or 100 uL of vehicle control dailiy from days 11 to 20 post aerosol M. tuberculosis challenge.

Techniques: Aerosol, Infection, Flow Cytometry, Control, Recombinant, MANN-WHITNEY

Journal: bioRxiv

Article Title: Mycobacterium tuberculosis suppresses protective Th17 responses during infection through multiple mechanisms

doi: 10.1101/2025.05.08.652811

Figure Lengend Snippet: Mice were infected with either the Erdman or Beijing strain of Mtb via the aerosol route and evaluated at 21dpi for (A) CFU in the lungs or (B-G) IL-17a + CD4 + T cells in the lungs. (B,C) unstimulated, (D,E) restimulated with Antigen 85B peptide, (F,G) restimulated with ESAT-6 peptide. Representative experiment of 2.

Article Snippet: Mice were intranasally administered 1 μg endotoxin-free recombinant murine IL-17a (R&D systems, 7956-ML-100/CF) in 40 μL sterile PBS daily or 40 μL of vehicle control daily, or intraperitoneally administered 2 μg endotoxin-free recombinant murine IL-17a in 100 μL sterile PBS daily or 100 uL of vehicle control dailiy from days 11 to 20 post aerosol M. tuberculosis challenge.

Techniques: Infection, Aerosol

Journal: bioRxiv

Article Title: Mycobacterium tuberculosis suppresses protective Th17 responses during infection through multiple mechanisms

doi: 10.1101/2025.05.08.652811

Figure Lengend Snippet: WT B6 mice were aerosol infected with indicated strains and evaluated at 21 days post infection. (A) Representative flow cytometry plots (cells gated on single, live, MHC-II − , Ly6G − , CD3 + , γδTCR − , CD4 + , CD8a − ) of IFN-γ + and IL-17a + CD4 + T cell populations. 21 days post infection, mice were sacrificed and evaluated ESAT-6 peptide pool stimulated lung homogenates for (B) IFN-γ + CD4 + T cells and (C) IL-17a + CD4 + T cells by flow cytometry. (E) Littermate-controlled WT B6 and Il17a −/− mice were aerosol infected with indicated strains at 21 days post infection with lung homogenate plated for CFU. Results are representative of 2 independent experiments. *, p < 0.05; **, p < 0.01 (unpaired nonparametric Mann-Whitney U test).

Article Snippet: Mice were intranasally administered 1 μg endotoxin-free recombinant murine IL-17a (R&D systems, 7956-ML-100/CF) in 40 μL sterile PBS daily or 40 μL of vehicle control daily, or intraperitoneally administered 2 μg endotoxin-free recombinant murine IL-17a in 100 μL sterile PBS daily or 100 uL of vehicle control dailiy from days 11 to 20 post aerosol M. tuberculosis challenge.

Techniques: Aerosol, Infection, Flow Cytometry, MANN-WHITNEY

Journal: bioRxiv

Article Title: Mycobacterium tuberculosis suppresses protective Th17 responses during infection through multiple mechanisms

doi: 10.1101/2025.05.08.652811

Figure Lengend Snippet: As in , WT B6 mice were aerosol infected with either WT, Δ eccC1, Δ eccC1::eccC1 , fadD28::tn , or fadD28::tn + p fadD28 M. tuberculosis Erdman strain. 21 days post infection, mice were sacrificed, lung single cell homogenates were stimulated with Antigen 85b peptide pool and measured for lung (A) IFN-γ + CD4 + T cells and (B) IL-17a + CD4 + T cells by flow cytometry. Results in A and B are representative of 2 independent experiments. *, p < 0.05; **, p < 0.01 (unpaired nonparametric Mann-Whitney U test

Article Snippet: Mice were intranasally administered 1 μg endotoxin-free recombinant murine IL-17a (R&D systems, 7956-ML-100/CF) in 40 μL sterile PBS daily or 40 μL of vehicle control daily, or intraperitoneally administered 2 μg endotoxin-free recombinant murine IL-17a in 100 μL sterile PBS daily or 100 uL of vehicle control dailiy from days 11 to 20 post aerosol M. tuberculosis challenge.

Techniques: Aerosol, Infection, Flow Cytometry, MANN-WHITNEY

Journal: bioRxiv

Article Title: Mycobacterium tuberculosis suppresses protective Th17 responses during infection through multiple mechanisms

doi: 10.1101/2025.05.08.652811

Figure Lengend Snippet: WT B6, Ifnar −/− , Sp140 −/− , Sp140 −/− Ifnar −/− , Ifng −/− , and Tbet −/− mice were aerosol infected with WT M. tuberculosis Erdman strain. (B, C) Representative flow cytometry plots (cells gated on single, live, MHC-II − , Ly6G − , CD3 + , γδTCR − , CD4 + , CD8a − ) of IFN-γ + and IL-17a + CD4 + T cells of these populations. 21 days post infection, lung homogenates were measured for (A) CFU and analyzed via flow cytometry for (D) IFN-γ + CD4 + T cells and (E) IL-17a + CD4 + T cells. Results in A, D, and E are representative of 2 independent experiments. *, p < 0.05; **, p < 0.01 (unpaired nonparametric Mann-Whitney U test).

Article Snippet: Mice were intranasally administered 1 μg endotoxin-free recombinant murine IL-17a (R&D systems, 7956-ML-100/CF) in 40 μL sterile PBS daily or 40 μL of vehicle control daily, or intraperitoneally administered 2 μg endotoxin-free recombinant murine IL-17a in 100 μL sterile PBS daily or 100 uL of vehicle control dailiy from days 11 to 20 post aerosol M. tuberculosis challenge.

Techniques: Aerosol, Infection, Flow Cytometry, MANN-WHITNEY

Journal: PLOS Pathogens

Article Title: Mucosal immunization with the lung Lactobacillus -derived amphiphilic exopolysaccharide adjuvanted recombinant vaccine improved protection against P . aeruginosa infection

doi: 10.1371/journal.ppat.1012696

Figure Lengend Snippet: (A) Experimental design and timeline of prime and booster regimen of PBS, rPcrV, CTB+rPcrV and EPS301@rPcrV vaccination (i.n.) and challenge. rPcrV-specific IgA titers in lung homogenate and IgG titers in serum were determined by ELISA before each round of immunization and every 14 days after the primary immunization. (B) The graph line shown the evolution of IgA titers in lung homogenate post immunization for each immunized group of mice. *p < 0.05; **p < 0.01; ***p < 0.001 for comparison with CTB+rPcrV immunized mice (n = 8). (C) The graph line shown the evolution of IgG titers in serum post immunization for each immunized group of mice. *p < 0.05 for comparison with rPcrV alone immunized mice (n = 8). (D) Mice were inoculated with 40 μl bacterial slurry (1×10 9 CFUs of P . aeruginosa PAO1) via left nostril on day 112 post booster vaccination and held upright for 1 min. Representative survival rates from two independent experiments are shown (n = 8–10). The data for survival test were analyzed by Wilcoxon log-rank survival test (*p < 0.05; **p < 0.01). (E) Mice were inoculated with 40 μl bacterial slurry (lower dose, 1×10 7 CFUs of P . aeruginosa PAO1) as above. At 12 h post challenge, the numbers of bacteria in lungs, BALF and spleens were counted (n = 5–8). (F) Mice were inoculated with the lower dose of PAO1 as above. At 12 h post challenge, histological evaluation of lung sections by light microscopy. Lung specimens were fixed, sectioned, and stained with H&E (n = 3–5). Data are presented as means ± SEM. Significant differences were calculated with One- or Two-way ANOVA followed by Tukey’s multiple comparisons test. ns, not significant, *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: To define the IL-17A and IFN-γ levels in lung and spleen tissue homogenate, the murine bioplex ELISA kits (R&D SYSTEMS Mouse IL-17A/F Heterodimer DuoSet ELISA DY5390 and R&D SYSTEMS Mouse IFN-gamma DuoSet ELISA DY485) were used according to the manufacturer’s recommendations.

Techniques: Enzyme-linked Immunosorbent Assay, Comparison, Bacteria, Light Microscopy, Staining

Journal: PLOS Pathogens

Article Title: Mucosal immunization with the lung Lactobacillus -derived amphiphilic exopolysaccharide adjuvanted recombinant vaccine improved protection against P . aeruginosa infection

doi: 10.1371/journal.ppat.1012696

Figure Lengend Snippet: Mice (n = 3-5/group) were immunized (i.n.) twice 14 days apart with rPcrV, CTB+ rPcrV or EPS301@rPcrV, with animals receiving PBS served as controls. Mice were inoculated with 40 μl bacterial slurry (lower dose, 1×10 7 CFUs of P . aeruginosa PAO1) as above. Vaccinated mice were sacrificed at 12 hours post-challenge on day 7 or day 112 after the second vaccination lung tissue were prepared. Number of IFN-γ + CD4 + T cells, IL-17A + CD4 + T cells (A), IFN-γ + γδ + T cells, IL-17A + γδ + T cells (B) in lung at 12 hours post-challenge on day 7 after the second vaccination were estimated by intracellular cytokine. Number of IFN-γ + CD4 + T cells, IL-17A + CD4 + T cells (C), IFN-γ + γδ + T cells, IL-17A + γδ + T cells (D) in lung at 12 hours post-challenge on day 112 after the second vaccination were estimated by intracellular cytokine. IFN-γ and IL-17 levels, determined by ELISA in a supernatant of lung tissue homogenate were analyzed. The IFN-γ levels and IL-17A levels (E) in lung at 12 hours post-challenge on day 7 and day 112 after the second vaccination were determined by ELISA. Data are presented as means ± SEM. Significant differences were calculated with One-way ANOVA followed by Tukey’s multiple comparisons test. *p < 0.05; **p < 0.01; ***p < 0.001 for comparison with EPS301@rPcrV immunized mice.

Article Snippet: To define the IL-17A and IFN-γ levels in lung and spleen tissue homogenate, the murine bioplex ELISA kits (R&D SYSTEMS Mouse IL-17A/F Heterodimer DuoSet ELISA DY5390 and R&D SYSTEMS Mouse IFN-gamma DuoSet ELISA DY485) were used according to the manufacturer’s recommendations.

Techniques: Enzyme-linked Immunosorbent Assay, Comparison

Journal: PLOS Pathogens

Article Title: Mucosal immunization with the lung Lactobacillus -derived amphiphilic exopolysaccharide adjuvanted recombinant vaccine improved protection against P . aeruginosa infection

doi: 10.1371/journal.ppat.1012696

Figure Lengend Snippet: (A) Comparative analysis of opsonophagocytosis against PAO1 using antisera from differently immunized mice. (B) Timeline of passive transfer and P . aeruginosa -induced pneumonia model. (C) Passive transfer of immunized and non-immunized mice was evaluated by i.v. injection of pooled serum (100 μl) to naïve C57BL/6 mice (n = 10/group). 24 hours after serum transfer, mice were inoculated with 40 μl bacterial slurry (1×10 9 CFUs of P . aeruginosa PAO1) via intranasal route (i.n.). Survival of mice from PBS, rPcrV, rPcrV+CTB and EPS301@rPcrV groups were observed for 48 h. The data for survival test were analyzed by Wilcoxon log-rank survival test (ns, not significant). (D) A lower dose of bacterial slurry (1×10 7 CFUs of P . aeruginosa PAO1) was inoculated to recipient mice via intranasal route (i.n.). Bacterial loads in lung and BALF were detected at 12 h post infection. (E) 12 hours post-challenge, lung tissue was prepared. IL-17A and IFN-γ levels determined by ELISA in a supernatant of lung tissue homogenate were analyzed. (F) Timeline of adoptive transfer and P . aeruginosa -induced pneumonia model. Lung and splenic CD3 + CD4 + T cells from EPS301@rPcrV–vaccinated congenic mice were purified on day 7 after second vaccination. CD3 + CD4 + T cells (5×10 4 ) were intravenously transferred into naïve C57BL/6 mice. (G) 24 hours after adoptive transfer, mice were inoculated with 40 μl bacterial slurry (1×10 9 CFUs of P . aeruginosa PAO1) via intranasal route (i.n.). Survival of mice were observed for 48 h. Survival after P . aeruginosa challenge (data were pooled from two independent experiments (n = 8–10). The data for survival test were analyzed by Wilcoxon log-rank survival test (***p < 0.001). (H) A lower dose of bacterial slurry (1×10 7 CFUs of P . aeruginosa PAO1) was inoculated to recipient mice via intranasal route (i.n.). Bacterial loads in lung and BALF were detected at 12 h post infection. (I) 12 hours post-challenge, lung tissue was prepared. IL-17A and IFN-γ levels determined by ELISA in a supernatant of lung tissue homogenate were analyzed. (J) Coexpression of CD44 and CD69 on non-stimulated lung CD4 + or γδ + T cells (gated in CD45 + CD4 + T cells). (K) FTY720 was initially dissolved in DMSO to create a 50 mg/mL stock solution. This stock solution was subsequently diluted with saline for intraperitoneal administration at a dosage of 30 μg per mice, and with distilled water for administration via stomach intubation at a dosage of 50 μg per mice. A lower dose of bacterial slurry (1×10 7 CFUs of P . aeruginosa PAO1) was inoculated to FTY720 treated and FTY720 non-treated mice via intranasal route (i.n.). Bacterial loads in lung were detected at 12 h post P . aeruginosa infection. Data are presented as means ± SEM. Significant differences were calculated with One-way ANOVA followed by Tukey’s multiple comparisons test (D, E, H and L), Mann-Whitney U test (J) or unpaired t test (K). *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: To define the IL-17A and IFN-γ levels in lung and spleen tissue homogenate, the murine bioplex ELISA kits (R&D SYSTEMS Mouse IL-17A/F Heterodimer DuoSet ELISA DY5390 and R&D SYSTEMS Mouse IFN-gamma DuoSet ELISA DY485) were used according to the manufacturer’s recommendations.

Techniques: Injection, Infection, Enzyme-linked Immunosorbent Assay, Adoptive Transfer Assay, Purification, Saline, MANN-WHITNEY

Journal: PLOS Pathogens

Article Title: Mucosal immunization with the lung Lactobacillus -derived amphiphilic exopolysaccharide adjuvanted recombinant vaccine improved protection against P . aeruginosa infection

doi: 10.1371/journal.ppat.1012696

Figure Lengend Snippet: TCR δ-deficient mice were vaccinated with EPS301@rPcrV and survival, bacterial loads and the lung pathology were detected on day 7 post second vaccination. Mice were inoculated with 40 μl bacterial slurry (1×10 9 CFUs of P . aeruginosa PAO1) via intranasal route (i.n.). (A) The number and percentage of Th 17 cells in WT and TCR δ-deficient mice before immunization, after immunization and post infection. (B) Survival of mice were observed for 48 h. Mice were inoculated with 40 μl bacterial slurry (1×10 7 CFUs of P . aeruginosa PAO1) via intranasal route (i.n.). The data for survival test were analyzed by Wilcoxon log-rank survival test (***p < 0.001). (C) Bacterial burdens of mice were counted at 12 h post infection. (D) Histological evaluation of lung sections by light microscopy. Lung specimens were fixed, sectioned, and stained with H&E (n = 3–5). (E) Timeline of adoptive transfer and P . aeruginosa -induced pneumonia model. Lung and splenic CD3 + γδ T cells from EPS301@rPcrV–vaccinated congenic mice were purified on day 7 after second vaccination. CD3 + γδ T cells (1 × 10 4 ) were intravenously transferred into naïve C57BL/6 mice. (F) 24 hours after adoptive transfer, mice were inoculated with 40 μl bacterial slurry (1×10 9 CFUs of P . aeruginosa PAO1) via intranasal route (i.n.). Survival of mice were observed for 48 h. Survival after P . aeruginosa challenge (data were pooled from two independent experiments (n = 8–10). The data for survival test were analyzed by Wilcoxon log-rank survival test (**p < 0.01, ***p < 0.001). A lower dose of bacterial slurry (1×10 7 CFUs of P . aeruginosa PAO1) was inoculated to recipient mice via i.n.. (G) Bacterial loads in lung and BALF were detected at 12 h post infection. (H) 12 hours post-challenge, lung tissue was prepared. IL-17A and IFN-γ levels determined by ELISA in a supernatant of lung tissue homogenate were analyzed. Data are presented as means ± SEM. Significant differences were calculated with unpaired t test (A) and One-way ANOVA followed by Tukey’s multiple comparisons test (C, G and H). ns, not significant, *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: To define the IL-17A and IFN-γ levels in lung and spleen tissue homogenate, the murine bioplex ELISA kits (R&D SYSTEMS Mouse IL-17A/F Heterodimer DuoSet ELISA DY5390 and R&D SYSTEMS Mouse IFN-gamma DuoSet ELISA DY485) were used according to the manufacturer’s recommendations.

Techniques: Infection, Light Microscopy, Staining, Adoptive Transfer Assay, Purification, Enzyme-linked Immunosorbent Assay

Journal: PLOS Pathogens

Article Title: Mucosal immunization with the lung Lactobacillus -derived amphiphilic exopolysaccharide adjuvanted recombinant vaccine improved protection against P . aeruginosa infection

doi: 10.1371/journal.ppat.1012696

Figure Lengend Snippet: (A) Experimental design and timeline of prime and booster regimen to IL-17A-deficient, IFN-γ-deficient and WT mice of EPS301-adjuvanted vaccination (i.n.) and challenge. (B) 7 days post booster vaccination, level of IgG in serum was detected by ELISA. (C) 7 days post booster vaccination, level of IgA in lung was detected by ELISA. (D) IL-17A-deficient, IFN-γ-deficient and WT mice were inoculated with 40 μl bacterial slurry (1×10 9 CFUs of P . aeruginosa PAO1) via left nostril on day 7 post booster vaccination and held upright for 1 min. Representative survival rates from two independent experiments are shown (n = 8–10). The data for survival test were analyzed by Wilcoxon log-rank survival test (**p < 0.01, ***p < 0.001). (E) Mice were inoculated with 40 μl bacterial slurry (lower dose: 1×10 7 CFUs of P . aeruginosa PAO1) as above. At 12 h, the numbers of bacteria in lung and BALF were counted (n = 5–8). (F) Mice were inoculated with the lower dose of PAO1 as above. At 12 h post challenge, histological evaluation of lung sections by light microscopy. Lung specimens were fixed, sectioned, and stained with H&E (n = 3–5). Data are presented as means ± SEM. Significant differences were calculated with One-way ANOVA followed by Tukey’s multiple comparisons test. ns, not significant, *p < 0.05, **p < 0.01, ***p < 0.001).

Article Snippet: To define the IL-17A and IFN-γ levels in lung and spleen tissue homogenate, the murine bioplex ELISA kits (R&D SYSTEMS Mouse IL-17A/F Heterodimer DuoSet ELISA DY5390 and R&D SYSTEMS Mouse IFN-gamma DuoSet ELISA DY485) were used according to the manufacturer’s recommendations.

Techniques: Enzyme-linked Immunosorbent Assay, Bacteria, Light Microscopy, Staining

Journal: iScience

Article Title: IL-22 and IL-23 regulate the anticryptococcal response during Cryptococcus deuterogattii infection

doi: 10.1016/j.isci.2024.111054

Figure Lengend Snippet:

Article Snippet: Murine IL-17A Standard ABTS ELISA Development Kit , PeproTech© , Cat#900-K382.

Techniques: Purification, Blocking Assay, Staining, Recombinant, Enzyme-linked Immunosorbent Assay, Activation Assay, Knock-Out, Software

Journal: International Journal of Molecular Sciences

Article Title: IL-17RA Signaling in Prx1+ Mesenchymal Cells Influences Fracture Healing in Mice

doi: 10.3390/ijms25073751

Figure Lengend Snippet: Conditional deletion of Il17ra in Prx1 + cells leads to subtle changes in bone microarchitecture. ( A ) Prx1-cre ; Il17ra F/F ( Il17ra cKO) were generated to conditionally delete Il17ra in Prx1 + mesenchymal cells by deleting exons 3 and 4 which leads to the early termination of translation. ( B ) Prx1 - cre significantly decreased the Il17ra gene expression in isolated periosteal cells. ( C ) Immunohistochemistry shows less immunoreactivity of IL-17RA within day 14 fracture calluses and the periosteum of Il17ra cKO mice. Scale bar: 50 µm. Representative 3D images of ( D ) femur trabecular bone, ( E ) femur cortical bone, and ( F ) L3 vertebral body trabecular bone. µCT analysis showing ( G ) femur trabecular bone volume fraction (BV/TV), ( H ) femur trabecular thickness (Tb.Th), ( I ) femur cortical thickness (Ct.Th), ( J ) femur cortical porosity (Ct.Po), ( K ) L3 vertebral body trabecular bone volume fraction (BV/TV), and ( L ) L3 vertebral body trabecular thickness (Tb.Th). Abbreviations: PO, periosteum; Prom, promoter; cKO, conditional knockout. Data represent mean ± SD. Student’s unpaired t -test, * p < 0.05, ** p < 0.001.

Article Snippet: To test the effects of IL-17a on fractured activated periosteal cells, murine IL-17A (Peprotech, Cranbury, NJ, USA; #210-17) was added to a differentiation medium at a final concentration of 20 and 50 ng/mL.

Techniques: Generated, Gene Expression, Isolation, Immunohistochemistry, Knock-Out

Journal: International Journal of Molecular Sciences

Article Title: IL-17RA Signaling in Prx1+ Mesenchymal Cells Influences Fracture Healing in Mice

doi: 10.3390/ijms25073751

Figure Lengend Snippet: Activation of IL-17RA signaling inhibits osteogenesis. ( A ) IL-17A at 20 and 50 ng/mL inhibited the gene expression of Runx2 , Osx , Cola1 , and Bglap in periosteal cells isolated from control mice. ( B ) IL-17A did not influence expression of Runx2 , Osx , Cola1 , and Bglap in Il 1 7ra cKO periosteal cells. ( C ) Alizarin red-S staining and quantification shows less mineralization in IL-17A-treated control periosteal cells at days 14 and 21 of osteogenic differentiation, but there was no effect on mineralization by Il17ra cKO cells. Dashed line represented a fold-change of 1. Abbreviations: Diff, osteogenic differentiation; Osx , osterix; Cola1 , collagen type 1; Bglap, osteocalcin. Data represent mean ± SD. The two-way ANOVA followed by Tukey’s multiple comparisons, * p < 0.05.

Article Snippet: To test the effects of IL-17a on fractured activated periosteal cells, murine IL-17A (Peprotech, Cranbury, NJ, USA; #210-17) was added to a differentiation medium at a final concentration of 20 and 50 ng/mL.

Techniques: Activation Assay, Gene Expression, Isolation, Control, Expressing, Staining

Journal: International Journal of Molecular Sciences

Article Title: IL-17RA Signaling in Prx1+ Mesenchymal Cells Influences Fracture Healing in Mice

doi: 10.3390/ijms25073751

Figure Lengend Snippet: IL-17RA signaling promotes periosteal progenitor cell migration. Wound healing assay shows that IL-17A (20 ng/mL) promoted cell migration at 12 h. ( Left ): representative images of scratched areas marked by black lines. ( Right ): the semi-quantitative analysis of wound closure was determined by measuring the widths of the wounds. Data represent the mean ± SD. Two-way ANOVA followed by Tukey’s multiple comparisons. Values not sharing a common letter differ significantly, p < 0.05.

Article Snippet: To test the effects of IL-17a on fractured activated periosteal cells, murine IL-17A (Peprotech, Cranbury, NJ, USA; #210-17) was added to a differentiation medium at a final concentration of 20 and 50 ng/mL.

Techniques: Migration, Wound Healing Assay