monoclonal rat anti il 33  (R&D Systems)

Journal: Molecular Psychiatry

Article Title: Requirement of brain interleukin33 for aquaporin4 expression in astrocytes and glymphatic drainage of abnormal tau

doi: 10.1038/s41380-020-00992-0

Figure Lengend Snippet: a , b Micrographs are reverted monochrome for FITC channel of AQP4 staining in the cerebral cortex a or hippocampus b in WT or Il33 −/− mice, revealing significant reduction of AQP4 after 60 weeks in Il33 −/− brains. CA , cornu ammonis; DG , dentate gyrus. c Statistical summary for immunofluorescence-based detection of AQP4 expression level demonstrates a significant reduction of AQP4 in both cerebral cortex and hippocampus at/after 60 weeks as compared with WT mice. IOD , integrated optical density. d Immunofluorescence shows disappearance of n-AQP4 in Il33 −/− brain at or after 60 weeks; note the presence of tubular/linear p-APQ4 (arrows) in Il33 −/− brains at 60 and 75 weeks; numerous age-related pigments in 75-week Il33 −/− brain are indicated by arrowheads. Silver staining (lower panels) demonstrates substantial numbers of neurites in Il33 −/− brain at 60 weeks, and nearly none at 75 weeks. e RT-PCR on cortical RNA shows a reduction of Aqp4 mRNA in Il33 −/− brains after 45 weeks. f Western blot of the cortical proteins shows reduction of AQP4 (green) in Il33 −/− mice at 65 weeks; α-actin (red) was used as internal control and probed simultaneously; its staining saturation is revealed (white) for more accurate evaluation. Bars’ unit = μm.

Article Snippet: Following antibodies were used in this study: biotin goat anti-mouse IL33 (R&D System, Minneapolis, MN, USA), monoclonal rat anti-mouse IL33 (clone 396118, ProSci, Charleston, SC, USA), rabbit anti-glial fibrillary acidic protein (GFAP) (astrocyte marker, Sigma-Aldrich, St. Louis, MO, USA), mouse monoclonal anti-tubulin β3 (neurons, R&D system), mouse monoclonal antibody AT8, which recognizes phosphor-tau(Ser202/Thr205) (ThermoFisher, Waltham, MA, USA), mouse monoclonal antibodies to PHF1 tau (Sigma-Aldrich), FITC-labeled or non-labeled anti-α-actin (Sigma-Aldrich), and rabbit anti-AQP4 antibody (Millipore).

Techniques: Staining, Immunofluorescence, Expressing, Silver Staining, Reverse Transcription Polymerase Chain Reaction, Western Blot, Control

Journal: Molecular Psychiatry

Article Title: Requirement of brain interleukin33 for aquaporin4 expression in astrocytes and glymphatic drainage of abnormal tau

doi: 10.1038/s41380-020-00992-0

Figure Lengend Snippet: a Immunofluorescence shows more PHF1 (green) in glomeruli of WT mice than Il33 −/− littermates at 70 weeks; right panels are enlarged boxed areas of left panels to show PHF1 in cytoplasm of glomerular mesangial cells (arrows). b Electron microscopy shows structural relationship of mesangial cells (MC) with other cells and glomerular basement membrane (GBM). c Western blot shows PHF1 in the cerebral cortex and isolated glomeruli of WT or Il33 −/− mice at 70 weeks; note an additional band of low molecular weight of PHF1 (arrow) among glomerular proteins, which is absent in cortical proteins. d RT-PCR shows a lack of tau mRNA in glomeruli as compared to the cortex in both WT and Il33 −/− mice. e Immunohistochemistry on PHF1 in the cerebral cortical and glomerular tissue from the same representative WT or Il33 −/− mice reveals a reverse correlation between PHF1 in the two tissue locations, i.e., more neuronal PHF1 vs less glomerular PHF1 (arrows) in Il33 −/− mice, and vice versa in WT mice. f Statistical analysis shows a positive regression between glomerular PHF1 and cortical AQP4 among WT and Il33 −/− mice. IOD , integrated optical density. g Immunofluorescence on normal human kidneys reveals the presence of PHF1, but not phospho-tau(Ser202/Thr205) (AT8), in glomeruli; an isotype IgG control is also shown; numbered boxed areas are enlarged to show PHF1 in mesangial cells. Bars’ unit = μm.

Article Snippet: Following antibodies were used in this study: biotin goat anti-mouse IL33 (R&D System, Minneapolis, MN, USA), monoclonal rat anti-mouse IL33 (clone 396118, ProSci, Charleston, SC, USA), rabbit anti-glial fibrillary acidic protein (GFAP) (astrocyte marker, Sigma-Aldrich, St. Louis, MO, USA), mouse monoclonal anti-tubulin β3 (neurons, R&D system), mouse monoclonal antibody AT8, which recognizes phosphor-tau(Ser202/Thr205) (ThermoFisher, Waltham, MA, USA), mouse monoclonal antibodies to PHF1 tau (Sigma-Aldrich), FITC-labeled or non-labeled anti-α-actin (Sigma-Aldrich), and rabbit anti-AQP4 antibody (Millipore).

Techniques: Immunofluorescence, Electron Microscopy, Membrane, Western Blot, Isolation, Molecular Weight, Reverse Transcription Polymerase Chain Reaction, Immunohistochemistry, Control

Journal: Molecular Psychiatry

Article Title: Requirement of brain interleukin33 for aquaporin4 expression in astrocytes and glymphatic drainage of abnormal tau

doi: 10.1038/s41380-020-00992-0

Figure Lengend Snippet: a Immunofluorescence shows a significant increase in p-AQP4 (red) expression in the cortex of Il33 −/− mice after receiving rIL33 injection ( Il33 −/− +rIL33) as compared with PBS controls ( Il33 −/− +PBS); two-color staining (lower panels) reveals high density of p-AQP4 (red) but nearly no granular n-APQ4 around neurons (green, tubulinβ3) in Il33 −/− +rIL33 mice, in contrast to many (arrows) in WT mice. Also note much less tubulinβ3 in both Il33 −/− +rIL33 and Il33 −/− +PBS mice than WT mice. b Immunofluorescence shows an increase in p-AQP4 expression in hippocampus (upper panels) of Il33 −/− +rIL33 mice; note that GFAP + astrocyte (green) population is similar among three groups (lower panel). c Western blot on cortical proteins shows quantity of AQP4 in Il33 −/− +rIL33 mice is comparable to WT mice, and much higher than Il33 −/− +PBS controls. d , e Statistics on total AQP4 expression (% of area) ( d ), and numbers of AQP4 + regions ( e ) in the cortex demonstrates that rIL33 injection enhances total AQP4 expression level but not numbers of AQP4 + regions.

Article Snippet: Following antibodies were used in this study: biotin goat anti-mouse IL33 (R&D System, Minneapolis, MN, USA), monoclonal rat anti-mouse IL33 (clone 396118, ProSci, Charleston, SC, USA), rabbit anti-glial fibrillary acidic protein (GFAP) (astrocyte marker, Sigma-Aldrich, St. Louis, MO, USA), mouse monoclonal anti-tubulin β3 (neurons, R&D system), mouse monoclonal antibody AT8, which recognizes phosphor-tau(Ser202/Thr205) (ThermoFisher, Waltham, MA, USA), mouse monoclonal antibodies to PHF1 tau (Sigma-Aldrich), FITC-labeled or non-labeled anti-α-actin (Sigma-Aldrich), and rabbit anti-AQP4 antibody (Millipore).

Techniques: Immunofluorescence, Expressing, Injection, Staining, Western Blot

Journal: Molecular Psychiatry

Article Title: Requirement of brain interleukin33 for aquaporin4 expression in astrocytes and glymphatic drainage of abnormal tau

doi: 10.1038/s41380-020-00992-0

Figure Lengend Snippet: a Immunohistochemistry shows neuronal PHF1 in indicated groups; note slightly reduced PHF1 in Il33 −/− +rIL33 mice as compared with Il33 −/− +PBS controls; no PHF1 was detectable in WT mice. Bars’ unit = μm b Regression analysis shows no linear correlation between total cortical AQP4 level (% of area) vs neuronal PHF1 among Il33 −/− +PBS, Il33 −/− +IL33, and WT mice; note that total AQP4 level in Il33 −/− +IL33 mice is not proportional to neuronal PHF1. Neuronal PHF1 was quantitated on digital immunostaining imagines, and expressed as IOD/neurons. c Statistical analysis shows a linear negative regression (black dotted line) between cortical and glomerular PHF1 among WT , Il33 −/− +PBS and Il33 −/− +rIL33 mice at 70 weeks; three mice were used for each group. d ELISA shows a sixfold increase in serum pCol peptide that had been injected into ventricles 24 h ago in Il33 −/− +rIL33 mice, as compared to PBS controls. e Diagram depicts two aqueous flows driven by astrocyte AQP4 (lightly colored arrows): Flow 1, driven by n-AQP4, brings neuronal wastes away from neurons, and Flow 2, convective flow driven by p-AQP4 toward perivenous space, flushes brain wastes further to perivenous space, i.e., glymphatics. Therefore, a lack of n-AQP4 results in ineffective removal of neuronal wastes, e.g., PHF1 from neurons.

Article Snippet: Following antibodies were used in this study: biotin goat anti-mouse IL33 (R&D System, Minneapolis, MN, USA), monoclonal rat anti-mouse IL33 (clone 396118, ProSci, Charleston, SC, USA), rabbit anti-glial fibrillary acidic protein (GFAP) (astrocyte marker, Sigma-Aldrich, St. Louis, MO, USA), mouse monoclonal anti-tubulin β3 (neurons, R&D system), mouse monoclonal antibody AT8, which recognizes phosphor-tau(Ser202/Thr205) (ThermoFisher, Waltham, MA, USA), mouse monoclonal antibodies to PHF1 tau (Sigma-Aldrich), FITC-labeled or non-labeled anti-α-actin (Sigma-Aldrich), and rabbit anti-AQP4 antibody (Millipore).

Techniques: Immunohistochemistry, Immunostaining, Enzyme-linked Immunosorbent Assay, Injection

Journal: Clinical and Experimental Immunology

Article Title: Experimentally induced psoriatic lesion associates with interleukin (IL)-6 in mast cells and appearance of dermal cells expressing IL-33 and IL-6 receptor

doi: 10.1111/j.1365-2249.2012.04618.x

Figure Lengend Snippet: Immunohistochemical staining of interleukin (IL)-33 in a 7-day-old psoriatic lesion (Köbner-positive skin). Note that the IL-33 staining is predominantly nuclear in the epidermis, but both nuclear and cytoplasmic stainings are seen in the dermis.

Article Snippet: Chemicals Mouse monoclonal anti-human IL-6 antibody (clone 1936), rat monoclonal anti-human IL-33 antibody (clone 390412) and recombinant human IL-6 (rh-IL-6) were purchased from R&D Systems Europe Ltd (Abingdon, UK), and a mouse monoclonal anti-IL-6R (CD126) antibody (catalogue number LS- {"type":"entrez-nucleotide","attrs":{"text":"C45194","term_id":"2381431","term_text":"C45194"}} C45194 ) from LifeSpan BioSciences (Seattle, WA, USA).

Techniques: Immunohistochemical staining, Staining

Journal: Neural Regeneration Research

Article Title: Gastrodin inhibits neuroinflammation in rotenone-induced Parkinson's disease model rats ★

doi: 10.3969/j.issn.1673-5374.2012.05.001

Figure Lengend Snippet: Area of interleukin-1β-positive cells (×10 3 μm 2 ) in the substantia nigra of rats

Article Snippet: IL-1β and IL-6 immunohistochemical staining: The primary antibodies were rabbit anti-rat IL-1β monoclonal antibody (1: 50; Wuhan Boster Bio-engineering Co., Ltd.) or rabbit anti-rat IL-6 monoclonal antibody (1: 50; Wuhan Boster Bio-engineering Co., Ltd.).

Techniques:

Journal: Neural Regeneration Research

Article Title: Gastrodin inhibits neuroinflammation in rotenone-induced Parkinson's disease model rats ★

doi: 10.3969/j.issn.1673-5374.2012.05.001

Figure Lengend Snippet: Interleukin-1β staining in the midbrains of rats (immunohistochemical staining, light microscopy; scale bars in A–E: 100 μm; scale bars in A’–E’: 500 μm). (A, A’) Normal group; (B, B’): Model group at 14 days; (C, C’): Model group at 28 days; (D, D’): Gastrodin group at 28 days; (E, E’): Madopar group at 28 days. Dark brown-colored cells were observed in the control (A), and the number of these was increased in the model at 14 days (B) and 28 days (C). The number of positive cells was reduced in the gastrodin (D) and Madopar groups at 28 days (E).

Article Snippet: IL-1β and IL-6 immunohistochemical staining: The primary antibodies were rabbit anti-rat IL-1β monoclonal antibody (1: 50; Wuhan Boster Bio-engineering Co., Ltd.) or rabbit anti-rat IL-6 monoclonal antibody (1: 50; Wuhan Boster Bio-engineering Co., Ltd.).

Techniques: Staining, Immunohistochemical staining, Light Microscopy, Control