Journal: Journal of Neuroinflammation

Article Title: Ceria nanoparticles ameliorate white matter injury after intracerebral hemorrhage: microglia-astrocyte involvement in remyelination

doi: 10.1186/s12974-021-02101-6

Figure Lengend Snippet: A1 astrocytes inhibited microglial phagocytosis of myelin debris via an astrocytic C3-microglial C3aR axis. a Representative images of GFAP and Iba1 double immunostaining in brain sections. Lens: × 100; Scale bar: 200 μm. b Representative TEM image showed microglia phagocytosis of myelin debris. Lens: × 7500, × 15,000; Scale bar: 5 μm (white), 2 μm (yellow). c tSNE plots of brain myeloid cells (Tabula Muris). d Expression of C3aR1 genes in microglia and macrophage (Tabula Muris). e Representative images of C3aR and Iba1 double immunostaining in brain sections. Lens: × 200; Scale bar: 50 μm. f Quantification of fluorescent beads in cultured microglia, * p < 0.01 versus Control, # p < 0.01 versus C3 treatment, ^ p < 0.01 versus A1 astrocyte CM treatment. g Representative images of fluorescent beads uptake in cultured microglia. Lens: × 400; Scale bar: 25 μm

Article Snippet: In the C3aR antagonist (C3aRA, SB290157, MCE HY-101502A) + A1 astrocyte CM group, 10 μM of C3aRA was incubated with microglia for 1 h before the A1 astrocyte CM treatment.

Techniques: Double Immunostaining, Expressing, Cell Culture, Control

Journal: Investigative ophthalmology & visual science

Article Title: Complement C3/C3aR Signaling Pathway Inhibition Ameliorates Retinal Damage in Experimental Retinal Vein Occlusion.

doi: 10.1167/iovs.66.5.2

Figure Lengend Snippet: FIGURE 4. C3aR antagonist downregulates expression of complement components C3 and C3aR in RVO. (A, B) RVO mice were intravitreally injected with vehicle or C3aR antagonist SB290157 (C3aRA) after RVO induction. Relative mRNA expression of C3 and C3ar in the control group (n = 4), the vehicle-treated group (n = 6), and the C3aRA-treated group (n = 4) RVO at 2 days. (C) Western blot analysis and quantification of C3 protein expression in retinal tissues (n = 8). (D) Representative immunofluorescence images showing IB4 (red) and C3 (green) staining in retinal flat-mount. (E) Quantification of C3 fluorescence intensity in the control, the vehicle-treated, and the C3aRA-treated RVO groups at 2 days (n = 4). One-way ANOVA with Tukey’s post hoc test. All data are presented as mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Then, 0.4% dimethyl sulfoxide (DMSO; 1 μL/eye) or the C3aR antagonist, SB290157 trifluoroacetate13 (C3aRA, 1 μg/μL/eye, Selleck Chemicals, USA, Cat. No: S8931) was administrated via intravitreal injection immediately after the RVO mouse model establishment.

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

Journal: JCI Insight

Article Title: Hypoxia-induced complement component 3 promotes aggressive tumor growth in the glioblastoma microenvironment

doi: 10.1172/jci.insight.179854

Figure Lengend Snippet: ( A ) C3AR1 expression of Pan-Cancer TCGA data of common cancer types ( n = 33). ( B ) C3AR1 expression in relation to glioma grade as analyzed in TCGA data. ( C ) C3AR1 expression in IDHwt glioma compared with IDHmut with or without 1p/19q codeletion (Tukey post hoc test) as analyzed in TCGA data. ( D ) C3AR1 expression in GBM compared with nontumor as analyzed in TCGA data. ( E ) Kaplan-Meier curve showing survival of patients with glioma with either high (red) or low (blue) C3AR1 expression based on TCGA data. ( F ) Kaplan-Meier curve showing survival of IDHwt GBM with high (red) or low (blue) C3AR1 expression based on TCGA data. ( G ) UMAP displaying C3AR1 expression in single-cell RNA-Seq data from 26 independent data sets compiled in GBmap . ( H ) C3AR1 expression of malignant cells divided into C3AR1 + (3.1%) or C3AR1 – (96.9%) cells. ( I ) GSEA of the C3AR1 -expressing malignant cells. Red bars indicate significant Benjamini-Hochberg adjusted P values ( P adj < 0.05). ( J ) Log-fraction plot of a combination of independent extreme limiting dilution sphere formation assays ( n = 4) of U3082MG glioma cells treated with SB290157. * P < 0.05, ** P < 0.01, or *** P < 0,001. One-way ANOVA ( B and C ), or unpaired t test ( D ) (in case of comparison between 2 groups) with Tukey post hoc test.

Article Snippet: For C3aR-antagonist studies, mice were randomized into groups and treated daily (5 days on, 2 days off) with 1 mg/kg SB290157 trifluoroacetate (Tocris, 6860/5) i.p., starting at the occurrence of glioma symptoms (day 37 ± 1) for up to a total of 30 injections.

Techniques: Expressing, RNA Sequencing, Comparison

Journal: JCI Insight

Article Title: Hypoxia-induced complement component 3 promotes aggressive tumor growth in the glioblastoma microenvironment

doi: 10.1172/jci.insight.179854

Figure Lengend Snippet: ( A and B ) Flow cytometry for indicated markers of M2 polarization in HMC3 microglia cells treated as indicated with SB290157 (SB), DMSO, and C3a ( n = 4). ( C ) Proliferation of HMC3 cells treated or not with SB290157 as measured by the WST-1 assay ( n = 3). ( D – F ) Flow cytometry for indicated markers of M2 (CD206, CD163) and M1 (CD86) polarization in primary murine macrophages treated as indicated with SB290157 (SB), DMSO, and C3a ( n = 3). ( G ) Representative image of immunofluorescence staining of CD31 and CD206 in murine gliomas induced through RCAS-PDGFB and RCAS-shp53 in Nestin/tv-a mice. Scale bars: 100 μm, 50 μm (insets). ( H ) Representative image of immunofluorescence staining of C3 and F4/80 in murine gliomas induced through RCAS-PDGFB and RCAS-shp53 in Nestin/tv-a mice. Scale bars: 50 μm.

Article Snippet: For C3aR-antagonist studies, mice were randomized into groups and treated daily (5 days on, 2 days off) with 1 mg/kg SB290157 trifluoroacetate (Tocris, 6860/5) i.p., starting at the occurrence of glioma symptoms (day 37 ± 1) for up to a total of 30 injections.

Techniques: Flow Cytometry, WST-1 Assay, Immunofluorescence, Staining

Journal: JCI Insight

Article Title: Hypoxia-induced complement component 3 promotes aggressive tumor growth in the glioblastoma microenvironment

doi: 10.1172/jci.insight.179854

Figure Lengend Snippet: ( A ) Illustration (created with BioRender.com) of study design for treatment with SB290157 (1 mg/kg) with or without combination with 10 Gy radiotherapy of murine gliomas induced through RCAS-PDGFB and RCAS-shp53 in Nestin/tv-a mice. ( B ) Kaplan-Meier curve showing the survival of mice treated with vehicle (red, n = 5), SB290157 (blue, n = 4), vehicle + 10 Gy (orange, n = 6), or SB290157 + 10 Gy (green, n = 8) as indicated. ( C ) Representative image of immunofluorescence staining of macrophages/microglia (F4/80), tumor cells (Olig2), and astrocytes (GFAP) in mice treated as indicated. ( D ) Quantitative analysis of F4/80 + cells/area (μm 2 ) as indicated ( n = 11 for untreated and n = 9 for SB290157 treated). ( E ) Representative images of immunofluorescence staining of tumor cells (Olig2) and CD206 + macrophages/microglia in mice treated as indicated. ( F ) Quantitative analysis of CD206 + cells/Area (μm 2 ) as indicated ( n = 11 for untreated and n = 9 for SB290157 treated). * P < 0.05, ** P < 0.01, or *** P < 0.001. Statistical analysis was performed using the Mann-Whitney U test. Scale bars: 100 μm.

Article Snippet: For C3aR-antagonist studies, mice were randomized into groups and treated daily (5 days on, 2 days off) with 1 mg/kg SB290157 trifluoroacetate (Tocris, 6860/5) i.p., starting at the occurrence of glioma symptoms (day 37 ± 1) for up to a total of 30 injections.

Techniques: Immunofluorescence, Staining, MANN-WHITNEY