Journal: Journal of Neuroinflammation

Article Title: Interleukin-1 receptor type 1 is overexpressed in neurons but not in glial cells within the rat superficial spinal dorsal horn in complete Freund adjuvant-induced inflammatory pain

doi: 10.1186/s12974-017-0902-x

Figure Lengend Snippet: Specificity of the anti-IL-1R1 antibody and distribution of IL-1R1 immunoreactivity in the spinal dorsal horn. a Adsorption of anti-IL-1R1 antibody to recombinant IL-1R1 peptide completely abolished the immunostaining. b Western blot analysis reinforces the specificity of the anti-IL-1R1 antibody. The single immunoreactive band indicates that the antibody detects a protein with a molecular mass of ~80 kDa that corresponds to the molecular weight of IL-1R1. c , d Micrographs showing immunoreactivity for IL-1R1 in control ( c ) and CFA-injected rats ( d ) 3 days after CFA-injection. Bars 100 μm

Article Snippet: As a part of the immunohistochemical protocol, we tested the specificity of the primary antibody on tissue sections by treating the diluted anti-IL-1R1 with recombinant rat IL-1R1 protein (Sino Biological Inc, Beijing, China, catalog no.: 80028R08H50) for the purpose of antibody adsorption.

Techniques: Adsorption, Recombinant, Immunostaining, Western Blot, Molecular Weight, Injection

Journal: Journal of Neuroinflammation

Article Title: Interleukin-1 receptor type 1 is overexpressed in neurons but not in glial cells within the rat superficial spinal dorsal horn in complete Freund adjuvant-induced inflammatory pain

doi: 10.1186/s12974-017-0902-x

Figure Lengend Snippet: CFA-evoked inflammation of the hindpaw initiates an overproduction of IL-1R1 protein in the spinal dorsal horn of rats. a Representative immune-blots showing immunoreactive bands for IL-1R1 and β-tubulin (loading control) in Western blots of tissue samples obtained from the L3–L5 lumbar segments of the spinal dorsal horn of the control and CFA-injected animals at post-injection day 3. b Histogram showing the optical densities of IL-1R1 immunostained bands (see on insert a ) calculated in proportion to the optical densities of β-tubulin (loading control) immunostained bands (see on insert a ). IL-1R1 protein level was found to be significantly higher than the control value ( p = 0.029). Data are shown as mean ± SEM

Article Snippet: As a part of the immunohistochemical protocol, we tested the specificity of the primary antibody on tissue sections by treating the diluted anti-IL-1R1 with recombinant rat IL-1R1 protein (Sino Biological Inc, Beijing, China, catalog no.: 80028R08H50) for the purpose of antibody adsorption.

Techniques: Western Blot, Injection

Journal: Journal of Neuroinflammation

Article Title: Interleukin-1 receptor type 1 is overexpressed in neurons but not in glial cells within the rat superficial spinal dorsal horn in complete Freund adjuvant-induced inflammatory pain

doi: 10.1186/s12974-017-0902-x

Figure Lengend Snippet: Mechanical withdrawal threshold and thermal withdrawal latency of wild type and IL-1R1 knockout mice during the course of CFA-induced inflammation of the hind paw. a The histogram shows the mechanical withdrawal threshold (MWT) of wild type (BL6) and IL-1R1 knockout (IL-1R1 KO) mice receiving different treatments in the three experimental groups: group (1) complete Freund-adjuvant (CFA) injection (BL6 CFA, IL-1R1 KO CFA); group (2) physiological saline injection (BL6 sham, IL-1R1 KO sham); and group (3) without any treatment (BL6 control, IL-1R1 KO control). Measurements were made only on the right hind paw which received the physiological saline or CFA injections. b The histogram shows the thermal withdrawal latency (TWL) of wild type (BL6) and IL-1R1 knockout (IL-1R1 KO) mice receiving different treatments in the three experimental groups: group (1) complet Freund-adjuvant (CFA) injection (BL6 CFA, IL-1R1 KO CFA); group (2) physiological salt solution injection (BL6 sham, IL-1R1 KO sham); and group (3) without any treatment (BL6 control, IL-1R1 KO control). Measurements were made only on the right hind paw which received the physiological saline or CFA injections. Data are shown as mean ± SEM

Article Snippet: As a part of the immunohistochemical protocol, we tested the specificity of the primary antibody on tissue sections by treating the diluted anti-IL-1R1 with recombinant rat IL-1R1 protein (Sino Biological Inc, Beijing, China, catalog no.: 80028R08H50) for the purpose of antibody adsorption.

Techniques: Knock-Out, Injection

Journal: Journal of Neuroinflammation

Article Title: Interleukin-1 receptor type 1 is overexpressed in neurons but not in glial cells within the rat superficial spinal dorsal horn in complete Freund adjuvant-induced inflammatory pain

doi: 10.1186/s12974-017-0902-x

Figure Lengend Snippet: Localization of IL-1R1 on neurons and glial cells in the superficial spinal dorsal horn of rats. Micrographs of single 1-μmthick laser scanning confocal optical sections illustrating the co-localization between immunolabeling for IL-1R1 ( red ; a – d , e , h , k , n , q ) and immunoreactivity for markers that are specific for axon terminals of peptidergic (CGRP, green ; a ) and non-peptidergic (IB4 binding, green ; b ) primary afferents, axon terminals of excitatory (VGLUT2, green ; c ) and inhibitory (VGAT, green ; d ) intrinsic neurons, astrocytes (GFAP, green ; o ) and microglial cells (CD11b, green ; r ), and postsynaptic membranes of excitarory (PSD95, green ; i ) and inhibitory (gephyrin, green ; l ) synapses in the superficial spinal dorsal horn. Mixed colors ( yellow ; marked by white arrowheads ) on the superimposed images ( j , m , p , s ) indicate double-labeled structures . The absence of yellow color on a – d indicates a lack of IL-1R1 expression on axon terminals of various origin. IL-1R1 immunoreactive spots appear in two different localization on the micrographs showing immunostaining also for KCC2 ( green , f , g ): (1) They can be aligned along the lines defined by the KCC2 immunostaining (cell membrane localization; white arrowheads on f and g ). (2) They can also be located in areas surrounded by the KCC2-immunostained cell membranes (cytoplasmic localization, yellow arrowhead on g . Bars 2 μm ( a – d ), 5 μm ( n – s ), and 10 μm ( e – m )

Article Snippet: As a part of the immunohistochemical protocol, we tested the specificity of the primary antibody on tissue sections by treating the diluted anti-IL-1R1 with recombinant rat IL-1R1 protein (Sino Biological Inc, Beijing, China, catalog no.: 80028R08H50) for the purpose of antibody adsorption.

Techniques: Immunolabeling, Binding Assay, Labeling, Expressing, Immunostaining

Journal: Journal of Neuroinflammation

Article Title: Interleukin-1 receptor type 1 is overexpressed in neurons but not in glial cells within the rat superficial spinal dorsal horn in complete Freund adjuvant-induced inflammatory pain

doi: 10.1186/s12974-017-0902-x

Figure Lengend Snippet: Histogram showing the CFA-evoked inflammation induced changes in the degree of co-localization between immunoreactivity for IL-1R1 and selected neuronal and glial markers in the superficial spinal dorsal horn of rats. Columns indicate the percentages of profiles immunoreactive for IL-1R1 that were found to be labeled also for the selected markers, and the ones that were aligned along KCC2 immunoreactive membranes (localization on the somatodendritic membrane of neurons) or were located within areas surrounded by KCC2 immunoreactive membranes (localization within the cytoplasm of the somatodendritic compartment of neurons). White columns show data obtained from control animals, whereas black columns represent values found in CFA-injected animals 3 days after CFA injection into the right hind paw. Asterisk indicate that CFA-evoked inflammation significantly increased the number of spots immunoreactive for IL-1R1 on the somato-denditic membrane of neurons ( p = 0.000001). Data are shown as mean ± SEM

Article Snippet: As a part of the immunohistochemical protocol, we tested the specificity of the primary antibody on tissue sections by treating the diluted anti-IL-1R1 with recombinant rat IL-1R1 protein (Sino Biological Inc, Beijing, China, catalog no.: 80028R08H50) for the purpose of antibody adsorption.

Techniques: Labeling, Injection

Journal: NPJ breast cancer

Article Title: IL-1B drives opposing responses in primary tumours and bone metastases; harnessing combination therapies to improve outcome in breast cancer.

doi: 10.1038/s41523-021-00305-w

Figure Lengend Snippet: Fig. 1 Depletion of IL-1B but not IL1R1 from the microenvironment promotes primary tumour growth. a Quantification (photons/sec (p/s)) of primary tumour growth in IL1R1fl/fl(n = 8 primary tumours from n = 4 mice) and IL1R1−/−(n = 8 primary tumours from n = 4 mice) mice up to 12 days of post-orthotopic injection of E0771-luc2-V5-GFP cells and b correspondent micrographs. c Quantification (p/s) of primary tumour growth in IL-1Bfl/fl(n = 7) and IL-1B−/−(n = 6) mice up to 26 days of post-orthotopic injection of E0771-luc2- GFP cells and d correspondent micrographs. IL-1Bfl/fl: n = 13 primary tumours from n = 7 mice (day 7 and 14); n = 12 primary tumours from n = 6 mice (day 20 and 26). IL-1B−/−: n = 12 primary tumours from n = 6 mice (day 7); n = 10 primary tumours from n = 6 mice (day 14, 20, and 26). 2.3-fold increase in primary tumour growth in IL-1B−/−mice (7.6 × 108 p/s) compared to IL-1Bfl/flmice (3.2 × 108 p/s) (P = 0.01). Data are mean +/−SEM, Two-way ANOVA with Sidak’s post-hoc test.

Article Snippet: The following plasmids were used for overexpression studies: IL-1B (MR226719L4, Origene), IL1R1 (MR227508L4, Origene), GFP (17448 pLenti CMV GFP Puro (658-5), Addgene), Luciferase (21474 pLenti CMV V5-LUC Blast (w567-1), Addgene).

Techniques: Injection

Journal: NPJ breast cancer

Article Title: IL-1B drives opposing responses in primary tumours and bone metastases; harnessing combination therapies to improve outcome in breast cancer.

doi: 10.1038/s41523-021-00305-w

Figure Lengend Snippet: Fig. 3 Tumour-derived IL-1B restores the infiltration of innate immune cells that may display anti-tumour functions and inhibits primary tumour growth in an IL-1B deficient microenvironment. a, b Images and quantification of primary tumour development in IL-1Bfl/fl(n = 8 primary tumours from n = 4 mice) and IL-1B−/−(n = 10 primary tumours from n = 5 mice) mice after intra-ductal administration of E0771 Luc2 V5 IL-1B-GFP cells. Data are mean +/−SEM, Two-way ANOVA with Sidak’s post-hoc test. c, d Quantification of F4/80+ macrophages (c) and CD163+ macrophages (d) in tumour core and periphery in IL-1Bfl/fland IL-1B−/−mice. Data are mean ± SEM, Two-way ANOVA with Sidak’s post hoc test. e, f Quantification of CD34+ blood vessels and MPO+ neutrophils in IL-1Bfl/fland IL-1B−/−mice. Data are shown as mean +/−SEM, Two-tailed unpaired t-test. g, h Images and quantification of primary tumour development in IL1R1fl/fl(n = 18 primary tumours from n = 9 mice) and IL1R1−/−(n = 14 primary tumours from n = 7 mice) mice after intra-ductal administration of E0771 Luc2 V5 IL-1B-GFP. Normalised data are shown as mean +/−SEM, Two-tailed unpaired t-test.

Article Snippet: The following plasmids were used for overexpression studies: IL-1B (MR226719L4, Origene), IL1R1 (MR227508L4, Origene), GFP (17448 pLenti CMV GFP Puro (658-5), Addgene), Luciferase (21474 pLenti CMV V5-LUC Blast (w567-1), Addgene).

Techniques: Derivative Assay, Two Tailed Test

Journal: NPJ breast cancer

Article Title: IL-1B drives opposing responses in primary tumours and bone metastases; harnessing combination therapies to improve outcome in breast cancer.

doi: 10.1038/s41523-021-00305-w

Figure Lengend Snippet: Fig. 4 IL-1B signalling enables breast cancer metastasis initiation. a, b Tumour proliferation after injection of E0771 luc2 V5 IL1R1-GFP cells in IL1R1fl/fl(n = 8 primary tumours from n = 4 mice) and IL1R1−/−(n = 6 primary tumours from n = 3 mice) mice. Data are mean +/−SEM, Two-way ANOVA with Sidak’s multiple comparisons test. c In vitro relative tumour growth of E0771 luc2 V5 GFP, IL-1B GFP or IL1R1 GFP cells upon stimulation with 40 pg/ml mouse recombinant IL-1B. Data are mean (+/−SEM), Two-way ANOVA with Sidak’s multiple comparison test (n = 6 technical repeats from two biological replicates). d Transwell cell migration of IL-1B overexpressing (P = 0.0009) and IL1R1 overexpressing (P < 0.0001) E0771 luc2 V5 cancer cells compared to control GFP-expressing cells (no treatment with exogenous IL-1B). Comparison of cell migration in vitro between E0771 luc2 V5 IL-1B GFP and IL1R1 GFP tumour cells (P = 0.0018). Data are mean (+/−SEM) cell number from 4 fields of view derived from three biological experiments (n = 12 technical repeats). Two-tailed unpaired t-test with Welch’s correction. e Representative images of haematoxylin-stained control, IL-1B and IL-1R1 overexpressing cells migrated through the membrane. Scale bar = 200 µm.

Article Snippet: The following plasmids were used for overexpression studies: IL-1B (MR226719L4, Origene), IL1R1 (MR227508L4, Origene), GFP (17448 pLenti CMV GFP Puro (658-5), Addgene), Luciferase (21474 pLenti CMV V5-LUC Blast (w567-1), Addgene).

Techniques: Injection, In Vitro, Recombinant, Comparison, Migration, Control, Expressing, Derivative Assay, Two Tailed Test, Staining, Membrane

Journal: Heliyon

Article Title: Pharmacodynamics of Sishen decoction in relieving rheumatoid arthritis: Chemical composition, regulatory pathway and online prediction simulation

doi: 10.1016/j.heliyon.2024.e37257

Figure Lengend Snippet: SSD treatment ameliorated RA rats. Representative images of hind paws from different groups and changes in foot circumference. Hematoxylin eosin staining and immunohistochemical staining (CD31, CD39, CD73, CCR6 and IL1R1) of knee joint synovial slices. * P < 0.05, ** P < 0.01 vs Model group.

Article Snippet: Additionally, antibodies for IL1R1, CD39, CD73, and CCR6 were obtained from Bosterbio in Wuhan, China.

Techniques: Staining, Immunohistochemical staining