Journal: Pharmaceutical biology

Article Title: Luteolin alleviated neutrophilic asthma by inhibiting IL-36γ secretion-mediated MAPK pathways.

doi: 10.1080/13880209.2022.2160770

Figure Lengend Snippet: Figure 3. Luteolin relieves airway inflammation and inhibits the activation of mitogen-activated protein kinase (MAPK) pathways in asthmatic mice. (A–D) IL-1b, phos- phorylated (p-)p38, p38, p-REK, ERK, p-JNK and JNK in mouse airway lung tissue as detected by western blotting. (E–H) Protein intensity analysis of IL-1b, p-p38, p38, p-REK, ERK, p-JNK and JNK in mouse airway lung tissue as detected by western blotting. p< 0.05 versus the control group. #p< 0.05 versus the asthma group.

Article Snippet: The glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and IL-1b polyclonal antibodies were purchased from Proteintech (Wuhan, China).

Techniques: Activation Assay, Western Blot, Control

Journal: Pharmaceutical biology

Article Title: Luteolin alleviated neutrophilic asthma by inhibiting IL-36γ secretion-mediated MAPK pathways.

doi: 10.1080/13880209.2022.2160770

Figure Lengend Snippet: Figure 4. Luteolin reduces the secretion of IL-36c and IL-1b in Beas-2B cells and inhibits the activation of mitogen-activated protein kinase (MAPK) pathways under lipopolysaccharide (LPS) stimulation. (A–D) IL-1b, p-p38, p38, p-REK, ERK, p-JNK and JNK in Beas-2B cells under LPS stimulation as detected by western blot. (E) IL-36c in Beas-2B cells under LPS stimulation as detected by enzyme-linked immunosorbent assay (ELISA). (F–I) Protein intensity analysis of IL-1b, p-p38, p38, p-REK, ERK, p-JNK and JNK in Beas-2B cells under LPS stimulation as detected by western blotting. p< 0.05 versus the control group. #p< 0.05 versus the LPS group.

Article Snippet: The glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and IL-1b polyclonal antibodies were purchased from Proteintech (Wuhan, China).

Techniques: Activation Assay, Western Blot, Enzyme-linked Immunosorbent Assay, Control

Journal: Pharmaceutical biology

Article Title: Luteolin alleviated neutrophilic asthma by inhibiting IL-36γ secretion-mediated MAPK pathways.

doi: 10.1080/13880209.2022.2160770

Figure Lengend Snippet: Figure 5. Luteolin reduces the secretion of IL-1b in Beas-2B cells and inhibits the activation of mitogen-activated protein kinase (MAPK) pathways under IL-36c stimu- lation. (A–D) IL-1b, p-p38, p38, p-REK, ERK, p-JNK and JNK in Beas-2B cells under IL-36c stimulation as detected by western blotting. (E–H) Protein intensity analysis of IL-1b, p-p38, p38, p-REK, ERK, p-JNK and JNK in Beas-2B cells under IL-36c stimulation as detected by western blotting. p< 0.05 versus the control group. #p< 0.05 versus the IL-36c group.

Article Snippet: The glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and IL-1b polyclonal antibodies were purchased from Proteintech (Wuhan, China).

Techniques: Activation Assay, Western Blot, Control

Journal: Pharmaceutical biology

Article Title: Luteolin alleviated neutrophilic asthma by inhibiting IL-36γ secretion-mediated MAPK pathways.

doi: 10.1080/13880209.2022.2160770

Figure Lengend Snippet: Figure 6. IL-36c siRNA reduces the secretion of IL-1b in Beas-2B cells and inhibits the activation of mitogen-activated protein kinase (MAPK) pathways under lipopoly- saccharide (LPS) stimulation. (A–D) IL-1b, p-p38, p38, p-REK, ERK, p-JNK, JNK in Beas-2B cells under LPS stimulation as detected by western blotting. (E–H) Protein intensity analysis of IL-1b, p-p38, p38, p-REK, ERK, p-JNK, JNK in Beas-2B cells under LPS stimulation as detected by western blotting. p< 0.05 versus the control group. #p< 0.05 versus the LPS þ NC siRNA group.

Article Snippet: The glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and IL-1b polyclonal antibodies were purchased from Proteintech (Wuhan, China).

Techniques: Activation Assay, Western Blot, Control

Journal: Pharmaceutical biology

Article Title: Luteolin alleviated neutrophilic asthma by inhibiting IL-36γ secretion-mediated MAPK pathways.

doi: 10.1080/13880209.2022.2160770

Figure Lengend Snippet: Figure 7. Schematic of the process by which luteolin reduces IL-1b by inhibiting the mitogen-activated protein kinase (MAPK) pathway via IL-36c.

Article Snippet: The glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and IL-1b polyclonal antibodies were purchased from Proteintech (Wuhan, China).

Techniques:

Journal: Stem Cells (Dayton, Ohio)

Article Title: The Vasoreparative Function of Myeloid Angiogenic Cells Is Impaired in Diabetes Through the Induction of IL1β

doi: 10.1002/stem.2810

Figure Lengend Snippet: MACs are proangiogenic cells with a distinct gene signature. (A): Venn diagram comparison between four datasets of M1 and M2 macrophage gene profiles. Heat maps of the common highly expressed M1 and M2 markers demonstrated that MACs gene signature is similar to M2 macrophages, whereas expression of M1 macrophage markers is weak/low. (B): qRT‐PCR data confirms MACs weak/low expression of M1 macrophage markers IL1β, ICAM1, IDO1, and PTGS2 in red; while MACs have a high expression of M2 macrophage markers CD163, CD206, CD204, and PDGFC in blue. n = 4; mean ± SEM. One‐way analysis of variance (ANOVA): p value not significant (ns), *, p < .05; **, p < .01; and ***, p < .001. (C): Cell surface immunophenotyping by flow cytometry for M1 macrophages, M2 macrophages, and MACs. Double stained isotype control was used for gating and shown as black contour plot. Red contour plot show M1 macrophages, blue contour plot show M2 macrophages, and green contour plot show MACs. Percentage is shown in each of four quadrants. n = 3. (D): Representative images of red‐labeled M1 macrophages, M2 macrophages, and MACs cocultured with green‐labeled ECFCs in 3D Matrigel Tubulogenesis assay. Scale bars: 500 µm. Quantification and statistical analysis of tube area. n = 3; mean ± SEM. One‐way ANOVA: p value not significant (ns), *, p < .05. Abbreviations: ECFCs, endothelial colony forming cells; MACs, myeloid angiogenic cells.

Article Snippet: Membranes were incubated with human IL1β antibody (MAB201, R&D Systems) or human Caspase‐1 antibody (MAB6215, R&D Systems) overnight at 4°C.

Techniques: Comparison, Expressing, Quantitative RT-PCR, Flow Cytometry, Staining, Control, Labeling

Journal: Stem Cells (Dayton, Ohio)

Article Title: The Vasoreparative Function of Myeloid Angiogenic Cells Is Impaired in Diabetes Through the Induction of IL1β

doi: 10.1002/stem.2810

Figure Lengend Snippet: HDG treatment shifts MACs toward a proinflammatory phenotype. (A): qRT‐PCR of MACs treated in HDG for 4 days resulted in an increase mRNA expression of inflammatory markers IL1β, IL6, and IL1α. Circles represent control, triangles HDG, and squares HLG. n = 3; mean ± SEM. One‐way analysis of variance (ANOVA): p value is not significant (ns), *, p < .05; **, p < .01; ***, p < .001. (B): Assessment of M2 macrophage markers CD163, CD204, and CD206 by qRT‐PCR in MACs treated with HDG for 4 days. n = 3; mean ± SEM. One‐way ANOVA: p value is not significant, *, p < .05; **, p < .01; and ***, p < .001. (C): Western blotting for pro‐IL1β and caspase1 in MAC whole cell lysates. MACs were exposed to HDG for 4 days and stimulated with LPS for 24 hours. RPL11 expression was used as the loading control. Representative images shown of n = 4. Abbreviations: C, control; HDG, high d ‐glucose; HLG, high l ‐glucose; MACs, myeloid angiogenic cells.

Article Snippet: Membranes were incubated with human IL1β antibody (MAB201, R&D Systems) or human Caspase‐1 antibody (MAB6215, R&D Systems) overnight at 4°C.

Techniques: Quantitative RT-PCR, Expressing, Control, Western Blot

Journal: Stem Cells (Dayton, Ohio)

Article Title: The Vasoreparative Function of Myeloid Angiogenic Cells Is Impaired in Diabetes Through the Induction of IL1β

doi: 10.1002/stem.2810

Figure Lengend Snippet: Inhibition of IL1β in high glucose conditions restores proangiogenic capacity of myeloid angiogenic cell (MAC)‐conditioned media (CM). (A): Representative images of Calcein green‐labeled endothelial colony forming cells grown in MAC‐CM in 3D Matrigel Tubulogenesis assay. Groups include: control; HDG; HLG; HDG plus addition of IL1β neutralizing antibody; addition of recombinant IL1β (rIL1β); HLG; HDG plus addition of normal mouse IgG. Scale bars: 500 µm. (B): Quantification and statistical analysis of tube area normalized to control shown as a box plot. n = 9–12; mean ± SEM. One‐way analysis of variance: p value is not significant versus control (ns in black), p value is not significant versus HDG (ns in green), **, p < .01 versus control, ***, p < .001 versus control, ### , p < .001 versus HDG. To facilitate data visualization, tube area ≥1 is shown in green, <1 in red, and osmotic control in blue. Abbreviations: HDG, high d ‐glucose; HLG, high l ‐glucose.

Article Snippet: Membranes were incubated with human IL1β antibody (MAB201, R&D Systems) or human Caspase‐1 antibody (MAB6215, R&D Systems) overnight at 4°C.

Techniques: Inhibition, Labeling, Control, Recombinant

Journal: Cancer letters

Article Title: NR0B2 re-educates myeloid immune cells to reduce regulatory T cell expansion and progression of breast and other solid tumors

doi: 10.1016/j.canlet.2024.217042

Figure Lengend Snippet: A cartoon overview of the inflammasome is depicted above the data, with the relevant part of the pathway highlighted in a box for reference. (A) Caspase 1 mRNA is decreased in BMDMs overexpressing NR0B2 (N = 3–4/group, t -test). (B) Caspase 1 mRNA is increased in CD11B + cells isolated from E0771 tumors grown in mice where NR0B2 was knocked out in myeloid immune cells (NR0B2 fl/fl ;LysMcre + ) (N = 4/group, t -test). (C) Intracellular caspase 1 activity was increased in BMDMs transfected an NR0B2 overexpression plasmid (N = 5/group, t -test). (D) Intracellular and secreted caspase 1 activity was increased in CD11B + cells isolated from E0771 tumors grown in mice where NR0B2 was knocked out in myeloid immune cells (NR0B2 fl/fl ;LysMcre + ) (N = 4/group, t -test). (E) Caspase 1 activity was increased in CD11C + cells isolated from E0771 tumors grown in mice where NR0B2 was knocked out in myeloid immune cells (NR0B2 fl/fl ;LysMcre + ) (N = 4/group, t -test). (F) IL1β mRNA is decreased in primary BMDMs transfected with an NR0B2 expression plasmid (N = 4/group, t -test). (G) IL1β mRNA is increased in primary BMDMs transfected with a shRNA plasmid against NR0B2 (N = 4/group, t -test). (H) IL1β mRNA is increased in CD11B + cells from primary E0771 tumors grown in mice where NR0B2 was knocked out in myeloid immune cells (NR0B2 fl/fl ;LysMcre + ) (N = 4/group, t -test). (I) IL1β mRNA is increased in CD11B + cells from E0771 lung metastases grown in mice where NR0B2 was knocked out in myeloid immune cells (NR0B2 fl/fl ;LysMcre + ) (N = 4/group, t -test). (J) IL1β protein secretion is decreased in BMDMs overexpressing NR0B2, but (K) increased in BMDMs from mice lacking NR0B2 in myeloid immune cells (N = 4/group, t -test). (L) Secreted IL1β protein is increased in splenic CD11C + cells isolated from mice where NR0B2 was knocked out in myeloid immune cells (NR0B2 fl/fl ;LysMcre + ) (N = 4/group, t -test). (M) Secreted IL1β protein is increased in CD11B + cells isolated from E0771 tumors grown in mice where NR0B2 was knocked out in myeloid immune cells (NR0B2 fl/fl ;LysMcre + ) (N = 4/group, t -test). For IL1β ELISA analysis, values below the detection limits were assigned a value of 0.

Article Snippet: To deplete IL1β (clone B122; BE0246; BioXCell) and IL18 (clone YIGIF74–1G7; BE0237; BioXCell), isotype control or respective neutralizing antibodies were added at 5 μg/mL to T cell cocultures.

Techniques: Isolation, Activity Assay, Transfection, Over Expression, Plasmid Preparation, Expressing, shRNA, Enzyme-linked Immunosorbent Assay

Journal: Cancer letters

Article Title: NR0B2 re-educates myeloid immune cells to reduce regulatory T cell expansion and progression of breast and other solid tumors

doi: 10.1016/j.canlet.2024.217042

Figure Lengend Snippet: (A) siRNA mediated knockdown of caspase 1 in BMDMs results in decreased T reg expansion, connecting the inflammasome within myeloid immune cells to T reg expansion. BMDMs were transfected with siRNA 24 h prior to co-culture with naive CD4 + T cells under Treg inducing conditions for additional 72hrs, T reg expansion was assessed by flow cytometry (N = 4/group, t -test). (B) siRNA against caspase 1 or IL1β attenuates induction of T regs in co-culture with myeloid cells from NR0B2 fl/fl ;LysMcre + mice. BMDMs were transfected with siRNA 24hrs prior to co-culture with naive CD4 + T cells under Treg inducing conditions for additional 72hrs, T reg expansion was assessed by flow cytometry (N = 5/group, 1-way ANOVA followed by Šidák’s multiple comparison test). (C) Immune neutralization with αIL1β attenuates induction of T regs in co-culture with BMDMs from NR0B2 fl/fl ;LysMcre + mice. BMDMs from control mice (NR0B2 +/+ ;LysMcre + ) or mice where NR0B2 was knocked out in myeloid immune cells (NR0B2 fl/fl ;LysMcre + ) were co-cultured with naïve CD4 + T cells under Treg inducing conditions in the presence of control αIgG or αIL1β neutralizing antibodies. 72hrs after co-culture, T reg expansion was assessed by flow cytometry (N = 5, 1-way ANOVA followed by Šidák’s multiple comparison test). (D) Antibody neutralization of IL1β attenuates the increased metastatic growth observed in mice lacking myeloid immune cell expression of NR0B2. Control NR0B2 +/+ ;LysMcre + or NR0B2 fl/fl ;LysMcre + mice were grafted intravenously with E0771 mammary cancer cells, and subsequently treated with a control IgG antibody or a neutralizing antibody against IL1β. Representative lung micrographs are shown. Red arrows denote lesions that were present on more than one consecutive serial section. (E) The DNA synthesis marker (proliferative index marker) Ki67 was measured as a proxy for metastatic burden in mice from (D). Ki67 mRNA expression, as measured by RT-qPCR, was increased in mice lacking myeloid immune cell expression of NR0B2, which was attenuated with αIL1β treatment (N = 9–15/group, 1-way ANOVA followed by Šidák’s multiple comparison test).

Article Snippet: To deplete IL1β (clone B122; BE0246; BioXCell) and IL18 (clone YIGIF74–1G7; BE0237; BioXCell), isotype control or respective neutralizing antibodies were added at 5 μg/mL to T cell cocultures.

Techniques: Knockdown, Transfection, Co-Culture Assay, Flow Cytometry, Comparison, Neutralization, Control, Cell Culture, Expressing, DNA Synthesis, Marker, Quantitative RT-PCR

Journal:

Article Title: Time of Day differences in the Number of Cytokine-, Neurotrophin- and NeuN-immunoreactive cells in the Rat Somatosensory or Visual Cortex

doi: 10.1016/j.brainres.2010.04.012

Figure Lengend Snippet: Quantitative evaluation of the number of TNF-, IL1β, NGF-, NeuN- and Fos-IR cells in the Sctx and Vctx layers at the end of the light (1100) or dark (2300) period. Data are represented as Mean ± S.E.M.

Article Snippet: Immunohistochemistry was performed as described previously ( Churchill et al., 2005 , 2008 ), using a polyclonal anti-human Fos antibody produced in rabbit (1:10,000; Calbiochem (Oncogene Research Products), San Diego, CA), goat anti-rat IL1β (0.5 μg/ml, R&D Systems, Inc., Minneapolis, MN), goat anti-rat TNFα (0.5 μg/ml, R&D Systems, Inc., Minneapolis, MN), rabbit anti-mouse NGF that cross reacts with rat NGF (1:2500 or 1:5000; Chemicon, Temecula, CA) or mouse monoclonal antibody to NeuN (1:500; Chemicon, Temecula, CA).

Techniques:

Journal:

Article Title: Time of Day differences in the Number of Cytokine-, Neurotrophin- and NeuN-immunoreactive cells in the Rat Somatosensory or Visual Cortex

doi: 10.1016/j.brainres.2010.04.012

Figure Lengend Snippet: IL1β-IR cells increased in Sctx layers II–V at 2300 h (C, D & F) in comparison to 1100 h (A, B, E). The IL1β-IR appears in the cytoplasm and extends into the apical dendrite of the large pyramidal neurons in layer V at 2300 h (D & F). Bar = 0.05 mm for A–D; Bar =0.025 mm for E&F.

Article Snippet: Immunohistochemistry was performed as described previously ( Churchill et al., 2005 , 2008 ), using a polyclonal anti-human Fos antibody produced in rabbit (1:10,000; Calbiochem (Oncogene Research Products), San Diego, CA), goat anti-rat IL1β (0.5 μg/ml, R&D Systems, Inc., Minneapolis, MN), goat anti-rat TNFα (0.5 μg/ml, R&D Systems, Inc., Minneapolis, MN), rabbit anti-mouse NGF that cross reacts with rat NGF (1:2500 or 1:5000; Chemicon, Temecula, CA) or mouse monoclonal antibody to NeuN (1:500; Chemicon, Temecula, CA).

Techniques: Comparison

Journal:

Article Title: Time of Day differences in the Number of Cytokine-, Neurotrophin- and NeuN-immunoreactive cells in the Rat Somatosensory or Visual Cortex

doi: 10.1016/j.brainres.2010.04.012

Figure Lengend Snippet: IL1β-IR cells increased in Vctx layers V–VI at 2300 h (D & F) in comparison to 1100 h (B, E). The IL1β-IR appears in the cytoplasm and extends into the apical dendrite of the large pyramidal neurons in layer V at 2300 h (D & F). Bar = 0.05 mm for A–D; Bar =0.025 mm for E&F.

Article Snippet: Immunohistochemistry was performed as described previously ( Churchill et al., 2005 , 2008 ), using a polyclonal anti-human Fos antibody produced in rabbit (1:10,000; Calbiochem (Oncogene Research Products), San Diego, CA), goat anti-rat IL1β (0.5 μg/ml, R&D Systems, Inc., Minneapolis, MN), goat anti-rat TNFα (0.5 μg/ml, R&D Systems, Inc., Minneapolis, MN), rabbit anti-mouse NGF that cross reacts with rat NGF (1:2500 or 1:5000; Chemicon, Temecula, CA) or mouse monoclonal antibody to NeuN (1:500; Chemicon, Temecula, CA).

Techniques: Comparison

Journal:

Article Title: Time of Day differences in the Number of Cytokine-, Neurotrophin- and NeuN-immunoreactive cells in the Rat Somatosensory or Visual Cortex

doi: 10.1016/j.brainres.2010.04.012

Figure Lengend Snippet: IL1β-IR astrocytes increased in number at 2300 h in layer VI (B& D) and the white matter below this layer (external capsule for the Sctx and forceps major (fmj) relative to 1100 h (A&C). The Vctx astrocytes (A&B) were much darker than the Sctx astrocytes (C & D). The layer VI and fibrous astrocytes in the white matter are marked by the arrows. Bar = 0.05 mm.

Article Snippet: Immunohistochemistry was performed as described previously ( Churchill et al., 2005 , 2008 ), using a polyclonal anti-human Fos antibody produced in rabbit (1:10,000; Calbiochem (Oncogene Research Products), San Diego, CA), goat anti-rat IL1β (0.5 μg/ml, R&D Systems, Inc., Minneapolis, MN), goat anti-rat TNFα (0.5 μg/ml, R&D Systems, Inc., Minneapolis, MN), rabbit anti-mouse NGF that cross reacts with rat NGF (1:2500 or 1:5000; Chemicon, Temecula, CA) or mouse monoclonal antibody to NeuN (1:500; Chemicon, Temecula, CA).

Techniques:

Journal:

Article Title: Time of Day differences in the Number of Cytokine-, Neurotrophin- and NeuN-immunoreactive cells in the Rat Somatosensory or Visual Cortex

doi: 10.1016/j.brainres.2010.04.012

Figure Lengend Snippet: Quantitative evaluation of the number of IL1β-IR astrocytes in the Sctx and Vctx layers at the end of the light (1100) or dark (2300) period. Data are represented as Mean ± S.E.M. for n= 7 rats.

Article Snippet: Immunohistochemistry was performed as described previously ( Churchill et al., 2005 , 2008 ), using a polyclonal anti-human Fos antibody produced in rabbit (1:10,000; Calbiochem (Oncogene Research Products), San Diego, CA), goat anti-rat IL1β (0.5 μg/ml, R&D Systems, Inc., Minneapolis, MN), goat anti-rat TNFα (0.5 μg/ml, R&D Systems, Inc., Minneapolis, MN), rabbit anti-mouse NGF that cross reacts with rat NGF (1:2500 or 1:5000; Chemicon, Temecula, CA) or mouse monoclonal antibody to NeuN (1:500; Chemicon, Temecula, CA).

Techniques: