Journal: Clinical and experimental immunology

Article Title: Targeting C3a/C5a receptors inhibits human mesangial cell proliferation and alleviates immunoglobulin A nephropathy in mice.

doi: 10.1111/cei.12961

Figure Lengend Snippet: Fig. 1. Cell proliferation and cytokine mRNA and protein expression in cultured human mesangial cells (HMCs). The HMCs were treated with blank medium or medium containing 100 lg/ml immunoglobulin (Ig)A, 100 lg/ml IgA 1 100 lM C3aR antagonist (C3aRA) or 100 lg/ml IgA 1 100 lM C5aR antagonist (C5aRA) for 48 h. The data are expressed as the mean 6 standard deviation (s.d.). (a) HMC proliferation was measured using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay (n 5 6 for each group). (b) Interleukin (IL)-6 (left row) and monocyte chemotactic protein 1 (MCP-1) (right row) gene expression levels relative to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were determined by reverse transcription–quantitative polymerase chain reaction (RT–qPCR). (c) Western blot analysis of IL-6, MCP-1 and GAPDH protein expression. (d) Quantitative analysis for Western blot of IL-6 and MCP-1 protein expression. *P < 005, **P < 001, ***P < 0001 versus negative control.

Article Snippet: The membranes were blocked with 5% skimmed milk solution in 0 1% Tris-buffered saline Tween-20 (Dingguo Bioscience) for 1 h. The membranes were incubated subsequently with either mouse antihuman C3aR monoclonal antibodies (1 : 500 dilution; AbD Serotec, Oxford, UK), mouse anti-human C5aR monoclonal antibodies (1 : 500 dilution; AbD Serotec), rabbit anti-human interleukin (IL)-6 monoclonal antibodies (1 : 800 dilution; Abcam, Cambridge, UK), rabbit antihuman MCP-1 polyclonal antibodies (1:1000 dilution, Abcam) or rabbit anti-human GAPDH polyclonal antibodies (1 : 800 dilution; Good Here Bioscience, Hangzhou, China) overnight at 4 C with gentle shaking.

Techniques: Expressing, Cell Culture, Standard Deviation, MTT Assay, Gene Expression, Reverse Transcription, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Western Blot, Negative Control

Journal: Clinical and experimental immunology

Article Title: Targeting C3a/C5a receptors inhibits human mesangial cell proliferation and alleviates immunoglobulin A nephropathy in mice.

doi: 10.1111/cei.12961

Figure Lengend Snippet: Fig. 2. Wild-type (WT), C3aR deficient (C3aR–/–) and C5aR–/– mice were immunized with inactivated Sendai virus to induce immunoglobulin (Ig)A nephropathy (IgAN). After 14 weeks of immunization, IgA and C3 deposition in the mesangium and the associated histological changes were analysed. (a,b) Glomerular IgA (upper row) and C3 (low row) deposition was measured by immunofluorescence staining of kidney sections from WT, C3aR–/– and C5aR–/– IgAN mice and negative controls. (c) Mesangial matrix expansion and cell proliferation were demonstrated by periodic acid-Schiff (PAS) staining of renal tissue in the following four groups of mice: negative controls and WT, C3aR–/– and C5aR–/– IgAN mice. Magnification 3400. Scale bars represent 100 lM.

Article Snippet: The membranes were blocked with 5% skimmed milk solution in 0 1% Tris-buffered saline Tween-20 (Dingguo Bioscience) for 1 h. The membranes were incubated subsequently with either mouse antihuman C3aR monoclonal antibodies (1 : 500 dilution; AbD Serotec, Oxford, UK), mouse anti-human C5aR monoclonal antibodies (1 : 500 dilution; AbD Serotec), rabbit anti-human interleukin (IL)-6 monoclonal antibodies (1 : 800 dilution; Abcam, Cambridge, UK), rabbit antihuman MCP-1 polyclonal antibodies (1:1000 dilution, Abcam) or rabbit anti-human GAPDH polyclonal antibodies (1 : 800 dilution; Good Here Bioscience, Hangzhou, China) overnight at 4 C with gentle shaking.

Techniques: Virus, Immunofluorescence, Staining

Journal: Clinical and experimental immunology

Article Title: Targeting C3a/C5a receptors inhibits human mesangial cell proliferation and alleviates immunoglobulin A nephropathy in mice.

doi: 10.1111/cei.12961

Figure Lengend Snippet: Fig. 3. Representative images of immunohistochemical staining for interleukin (IL)-6 and monocyte chemotactic protein 1 (MCP-1) in the kidney sections of wild-type (WT), C3aR deficient (C3aR–/–) and C5aR–/– immunoglobulin (Ig)A nephropathy (IgAN) mice and negative controls. C3aR and C5aR deficiency reduced renal IL-6 and MCP-1 expression compared to WT mice. Scale bars represent 100 lM.

Article Snippet: The membranes were blocked with 5% skimmed milk solution in 0 1% Tris-buffered saline Tween-20 (Dingguo Bioscience) for 1 h. The membranes were incubated subsequently with either mouse antihuman C3aR monoclonal antibodies (1 : 500 dilution; AbD Serotec, Oxford, UK), mouse anti-human C5aR monoclonal antibodies (1 : 500 dilution; AbD Serotec), rabbit anti-human interleukin (IL)-6 monoclonal antibodies (1 : 800 dilution; Abcam, Cambridge, UK), rabbit antihuman MCP-1 polyclonal antibodies (1:1000 dilution, Abcam) or rabbit anti-human GAPDH polyclonal antibodies (1 : 800 dilution; Good Here Bioscience, Hangzhou, China) overnight at 4 C with gentle shaking.

Techniques: Immunohistochemical staining, Staining, Expressing

Journal: Clinical and experimental immunology

Article Title: Targeting C3a/C5a receptors inhibits human mesangial cell proliferation and alleviates immunoglobulin A nephropathy in mice.

doi: 10.1111/cei.12961

Figure Lengend Snippet: Fig. 4. Cytokine and chemokine gene expression in the renal tissues of the following four groups of mice: negative controls and wild-type (WT), C3aR deficient (C3aR–/–) and C5aR–/– immunoglobulin (Ig)A nephropathy (IgAN) mice (n5 7 for each group). Reverse transcription–quantitative polymerase chain reaction (RT–qPCR) was used to quantify the mRNA expression levels of (a) tumour necrosis factor (TNF)-a, (b) transforming growth factor (TGF)-b, (c) interleukin (IL)-1b, (d) interleukin (IL)-6 and (e) monocyte chemotactic protein 1 (MCP-1) relative to glyceraldehyde-3- phosphate dehydrogenase (GAPDH). Data are expressed as the mean 6 standard deviation. *P< 005, **P< 001, ***P< 0001 versus negative control.

Article Snippet: The membranes were blocked with 5% skimmed milk solution in 0 1% Tris-buffered saline Tween-20 (Dingguo Bioscience) for 1 h. The membranes were incubated subsequently with either mouse antihuman C3aR monoclonal antibodies (1 : 500 dilution; AbD Serotec, Oxford, UK), mouse anti-human C5aR monoclonal antibodies (1 : 500 dilution; AbD Serotec), rabbit anti-human interleukin (IL)-6 monoclonal antibodies (1 : 800 dilution; Abcam, Cambridge, UK), rabbit antihuman MCP-1 polyclonal antibodies (1:1000 dilution, Abcam) or rabbit anti-human GAPDH polyclonal antibodies (1 : 800 dilution; Good Here Bioscience, Hangzhou, China) overnight at 4 C with gentle shaking.

Techniques: Gene Expression, Reverse Transcription, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Expressing, Standard Deviation, Negative Control

Journal: BMC Genomics

Article Title: Transcriptional profiling reveals developmental relationship and distinct biological functions of CD16+ and CD16- monocyte subsets

doi: 10.1186/1471-2164-10-403

Figure Lengend Snippet: Real-time RT-PCR validation of microarray results . The expression of CD16, C3AR1, C1QR1, ICAM-2, CSFR1, CSF3R, CDKN1C, TNFRSF8, and LTB mRNA was quantified by SYBR Green real time RT-PCR in CD16 + and CD16 - Mo. The concentration of each gene was normalized to the 28S rRNA internal control and expressed as fgs RNA of a target gene per 1 ng rRNA28S. Depicted are results (mean ± SD of triplicate wells; *, p < 0.05, unpaired t-test, CD16 + versus CD16 - Mo) obtained with matched cells from 2 different healthy donors.

Article Snippet: Fluorochrome-conjugated Abs used for FACS analysis were CD14, CD16, CD19, CD16b, CD66b, CD56, and CD3 (Beckman Coulter); M-DC8 and CD1c (Miltenyi), HLA-DR, CD114, C3aR, CD1d and CD43 (BD Pharmingen), CD115 (R&D Systems), CD93/C1qR1 (Chemicon International), and CXCL16 (R&D Systems).

Techniques: Quantitative RT-PCR, Biomarker Discovery, Microarray, Expressing, SYBR Green Assay, Concentration Assay, Control

Journal: BMC Genomics

Article Title: Transcriptional profiling reveals developmental relationship and distinct biological functions of CD16+ and CD16- monocyte subsets

doi: 10.1186/1471-2164-10-403

Figure Lengend Snippet: Differential expression of CD114/CSF3R, CD115/CSF1R, CD93/C1qR1 and C3aR1 on CD16 + and CD16 - monocytes . Freshly isolated PBMC were stained with FITC CD14, PE-Cy5 CD16, and PE CD114, PE CD115, and PE CD93 Abs. The expression of CD3aR1 was detected after staining with unconjugated mouse C3AR1 Ab and PE rat anti-mouse Ab (RAM). CD14 high CD16 neg (R2) and CD14 low CD16 + (R3) Mo (A) were analyzed for expression of CD114, CD115, CD93 and C3aR1 (B) . Shown is an overlay histogram from one representative donor of 4 donors examined (B, left panels) and graphs showing mean ± SEM for % or MFI of CD114, CD115, CD93 and C3aR1 expression on each Mo subset (B, right panels) . (*, Paired t-test p-value < 0.05, CD16 + versus CD16 - Mo; n = 4).

Article Snippet: Fluorochrome-conjugated Abs used for FACS analysis were CD14, CD16, CD19, CD16b, CD66b, CD56, and CD3 (Beckman Coulter); M-DC8 and CD1c (Miltenyi), HLA-DR, CD114, C3aR, CD1d and CD43 (BD Pharmingen), CD115 (R&D Systems), CD93/C1qR1 (Chemicon International), and CXCL16 (R&D Systems).

Techniques: Quantitative Proteomics, Isolation, Staining, Expressing

Journal: bioRxiv

Article Title: Hypoxia-induced Complement Component 3 Promotes Aggressive Tumor Growth in the Glioblastoma Microenvironment

doi: 10.1101/2024.01.28.577617

Figure Lengend Snippet: A-B. Immunofluorescent detection of C3 (red), Nestin (yellow), Nuclei (DAPI, blue), GFAP (green) or Olig2 (cyane) in the perivascular (A) or hypoxic (B) niches of murine RCAS-PDGFB- and RCAS-shp53-induced gliomas. C-D. Hallmark_Hypoxia (C) and Hallmark_Complement (D) signatures mapped on spatially resolved transcriptomics from human GBM samples, displayed with Z-score. E. Spatial correlation between Hallmark_Hypoxia and Hallmark_Complement in one representative tumor (UKF242). P values were corrected for multiple hypothesis testing using the Holm method. F. Distribution of R-values for the correlation between Hallmark_Hypoxia and Hallmark_Complement in a total of 19 human GBM tissue samples with an average correlation score of R=0.54 G-H. Pearson correlation coefficient between Hallmark_Complement and Hallmark_Hypoxia signatures in the TCGA GBMLGG (G) or TCGA GBM (H) dataset. I-K. Expression of CA9, C3, and C3AR1 mRNA in human primary astrocytes (n=3), HMC3 microglia (n=4), or primary human glioma cells U3082MG (n=3), U3065MG (n=3) or U3084MG (n=3) in response to normoxia (21% O2), hypoxia (1% O2) or severe hypoxia (0.1% O2). Error bars represent SEM. *, P < 0.05, **, P < 0.01, or ***, P< 0,001. Statistical tests were one-way ANOVA, with Tukey post-hoc test, or unpaired t-test in case of two sample comparisons.

Article Snippet: The spheres were grown until visible spheres were formed (up to 14 days) with control or treatments with 50 and 250nM C3aR antagonist (Santa Cruz Biotechnology, SB290157).

Techniques: Expressing

Journal: bioRxiv

Article Title: Hypoxia-induced Complement Component 3 Promotes Aggressive Tumor Growth in the Glioblastoma Microenvironment

doi: 10.1101/2024.01.28.577617

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 IDH wildtype glioma compared to IDH mutant with or without 1p/19q codeletion (Tukey post-hoc test) as analyzed in TCGA data. D. C3AR1 expression in GBM compared to non-tumor as analyzed in TCGA data. E. Kaplan-Meier curve showing survival of glioma patients with either high (red) or low (blue) C3AR1 expression based on TCGA data. F . Kaplan-Meier curve showing survival of IDH-wildtype GBM with high (red) or low (blue) C3AR1 expression based on TCGA data. G. UMAP displaying C3AR1 expression in single cell sequencing data from 26 independent datasets compiled in GBmap . H. C3AR1 expression of malignant cells divided into C3AR1-expressing or non-expressing cells. I. Geneset enrichment analysis of the C3AR1-expressing malignant cells. Red colored bars indicate significant Benjamini-Hochberg adjusted P values (padj < 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, or unpaired T-test (in case of comparison between two groups) with Tukey post-hoc test.

Article Snippet: The spheres were grown until visible spheres were formed (up to 14 days) with control or treatments with 50 and 250nM C3aR antagonist (Santa Cruz Biotechnology, SB290157).

Techniques: Expressing, Mutagenesis, Sequencing, Comparison

Journal: Computational and Structural Biotechnology Journal

Article Title: The complement C3a/C3aR pathway is associated with treatment resistance to gemcitabine-based neoadjuvant therapy in pancreatic cancer

doi: 10.1016/j.csbj.2024.09.032

Figure Lengend Snippet: Complement C3a is highly expressed and is associated with neoadjuvant chemotherapy resistance in pancreatic cancer. (A) Venn diagram, including 3 parts of data: (1) differential genes associated with distinct gemcitabine response (in purple color); (2) differential genes associated with distinct immune infiltration (in blue color); (3) differential genes associated with distinct IC50 value of gemcitabine (in gray color). C3aR1 was the only differential gene shared by all these three parts. (B) Analysis of clinical samples from TCGA(PAAD) database showed that high expression of C3aR1 was associated with poor survival in pancreatic cancer. (C) The boxplot showing the differential expression of C3aR1 in pancreatic cancer tissue (T) and normal pancreatic tissue (N). (D) Representative IHC staining images showed the differential expression of C3, C3a, and C3aR in normal pancreatic tissue and human pancreatic cancer. (E) Representative images of H&E staining and IHC staining showed the differential expression of C3a, and C3aR in normal pancreatic tissue from wild-type (WT) C57BL/6 J mice and in pancreatic cancer tissue from KPC mice. (F) C3a protein levels in human pancreatic cancer tissue and adjacent normal pancreatic tissue were accessed by ELISA. (G) Representative IHC staining images of high/low C3aR expression in pancreatic cancer tissue from FUSCC-TMA. (H) Kaplan–Meier survival analysis of overall survival between high and low C3aR groups in FUSCC-TMA samples. (I) Representative IHC staining images showed the differential expression of C3a and C3aR in pancreatic cancer tissue from gemcitabine-sensitive (GS) patients and gemcitabine-resistant patients. (J) C3a protein levels in pancreatic cancer tissue from GS/GR patients were accessed by ELISA. * P < 0.05, * * P < 0.01, * ** P < 0.001. GEM, gemcitabine.

Article Snippet: Recombinant human complement C3a protein (MedChemExpress, MCE; HY-P7862), recombinant mouse complement C3a protein (MCE; HY-P7863) and C3aR antagonist, SB290157 (TargetMol) were used according to the manufacturer’s instructions.

Techniques: Expressing, Quantitative Proteomics, Immunohistochemistry, Staining, Enzyme-linked Immunosorbent Assay

Journal: Computational and Structural Biotechnology Journal

Article Title: The complement C3a/C3aR pathway is associated with treatment resistance to gemcitabine-based neoadjuvant therapy in pancreatic cancer

doi: 10.1016/j.csbj.2024.09.032

Figure Lengend Snippet: The activation of C3a/C3aR pathway promotes the proliferation, migration and gemcitabine resistance in pancreatic cancer cells. (A) Western blotting was used to detect the C3aR expression in Panc-1 cells treated with PBS or 500 ng/ml recombinant C3a protein. (B) Representative images of EdU assays showed that C3a recombinant protein promotes the proliferation of Panc-1 cells in a concentration-dependent manner. (C) The bar plot showed the percentage of EdU + cells per field. (D) Representative images of Transwell assays showed that C3a recombinant protein promotes the migration of Panc-1 cells in a concentration-dependent manner. (E) The bar plot showed the percentage of migrated cells per field. (F) CCK-8 assay showed that C3a recombinant protein increased the IC50 values of Panc-1 cells to gemcitabine. (G) Western blotting was used to detect the C3aR expression in Panc-02 cells treated with PBS or 500 ng/ml recombinant C3a protein (mouse). (H) Representative images of EdU assays showing that C3a recombinant protein (mouse) promotes the proliferation of Panc-02 cells in a concentration-dependent manner. (I) The bar plot showed the percentage of EdU + cells per field. (J) Representative images of Transwell assays showing that C3a recombinant protein (mouse) promotes the migration of Panc-02 cells in a concentration-dependent manner. (K) The bar plot showed the percentage of migrated cells per field. (L) CCK-8 assay showed that C3a recombinant protein (mouse) increased the IC50 values of Panc-02 cells to gemcitabine. * P < 0.05, * * P < 0.01, * ** P < 0.001. GEM, gemcitabine.

Article Snippet: Recombinant human complement C3a protein (MedChemExpress, MCE; HY-P7862), recombinant mouse complement C3a protein (MCE; HY-P7863) and C3aR antagonist, SB290157 (TargetMol) were used according to the manufacturer’s instructions.

Techniques: Activation Assay, Migration, Western Blot, Expressing, Recombinant, Concentration Assay, CCK-8 Assay

Journal: Computational and Structural Biotechnology Journal

Article Title: The complement C3a/C3aR pathway is associated with treatment resistance to gemcitabine-based neoadjuvant therapy in pancreatic cancer

doi: 10.1016/j.csbj.2024.09.032

Figure Lengend Snippet: Knockdown of C3aR does not affect the progression of pancreatic cancer cells in vitro. A.A-F Panc-1 cells were infected with C3aR shRNA to knock down the C3aR expression (sh#1, sh#2), or infected with a scrambled shRNA as the control group (sh-Scr). (A) Western blotting was used to detect C3aR expression in control cells and C3aR-deficient cells to evaluate the transfection efficiency. (B) CCK8 assay was used to evaluate the cell proliferation, and the relative optical density of each group at day 0, 1, 2, 3 and 4 was measured. The line chart showed that C3aR knockdown did not change the proliferation capacity of Panc-1 cells. (C) Representative images of EdU assays showed that the knockdown of C3aR did not affect the proliferation capacity of Panc-1 cells. (D) The bar plot showed the percentage of EdU + cells per field. (E) Representative images of Transwell assays showed that the knockdown of C3aR did not affect the migration ability of Panc-1 cells. (F) The bar plot showed the percentage of migrated cells per field. G-K C3aR-deficient Panc-1cells or control cells were treated with C3a recombinant protein at a concentration of 0 or 500 ng/ml. (G) RT-qPCR was used to detect the C3aR expression in control cells and C3aR-deficient cells with or without C3a treatment. (H) Representative images of EdU assays showed that the proliferation ability of control cells enhanced under the C3a treatment, while that of C3aR-deficient Panc-1 cells did not change. (I) The bar plot showed the percentage of EdU + cells per field. (J) Representative images of Transwell assays showed that the migration ability of control cells enhanced under the C3a treatment, while that of C3aR-deficient Panc-1 cells did not change. (K) The bar plot showed the percentage of migrated cells per field. L-O Ten female BALB/c-nu mice were randomly divided into 2 groups and respectively injected with control Panc-1 cells and C3aR-deficient cells to construct subcutaneous tumor model. Tumor volume was observed and recorded every 3 days. (L) Line charts of volume changes of subcutaneous tumor. (M) On the 21st of treatment, subcutaneous tumors were separated to show tumor size. (N) Western blotting was used to detect the expression of C3aR in subcutaneous tumors. (O) Elisa was used to detect the expression of complement C3a in subcutaneous tumors. (P) Frozen sections were prepared from tumor tissues. IF staining with antibodies to Ki-67 (in red color) was performed to detect proliferation of Panc-1 cells, and DAPI was used to counterstain the cell nuclei (in blue color). (Q) The bar plot showed the percentage of Ki-67 + cells per field.

Article Snippet: Recombinant human complement C3a protein (MedChemExpress, MCE; HY-P7862), recombinant mouse complement C3a protein (MCE; HY-P7863) and C3aR antagonist, SB290157 (TargetMol) were used according to the manufacturer’s instructions.

Techniques: Knockdown, In Vitro, Infection, shRNA, Expressing, Control, Western Blot, Transfection, CCK-8 Assay, Migration, Recombinant, Concentration Assay, Quantitative RT-PCR, Injection, Construct, Enzyme-linked Immunosorbent Assay, Staining

Journal: Computational and Structural Biotechnology Journal

Article Title: The complement C3a/C3aR pathway is associated with treatment resistance to gemcitabine-based neoadjuvant therapy in pancreatic cancer

doi: 10.1016/j.csbj.2024.09.032

Figure Lengend Snippet: C3aR antagonist (SB290157) attenuated the effects of C3a activation in pancreatic cancer. (A) Representative images of EdU assays showed that the treatment of SB290157 attenuated C3a-induced proliferation in Panc-1 cells. (B) The bar plot showed the percentage of EdU + cells per field. (C) Representative images of Transwell assays showed that the treatment of SB290157 attenuated C3a-induced migration in Panc-1 cells. (D) The bar plot showed the percentage of migrated cells per field. (E) CCK-8 assay showed that the treatment of SB290157 attenuated C3a-induced gemcitabine resistance in Panc-1 cells. F-K A mouse subcutaneous tumor model of Panc-1 cells was constructed. The mice were randomly grouped and treated with vehicle, 20 mg/kg gemcitabine, 20 mg/kg SB290157, 20 mg/kg gemcitabine combined with 20 mg/kg SB29015 by intraperitoneal injection once a day. (F) Line charts of volume changes of subcutaneous tumor. (G) On the 14th day of treatment, subcutaneous tumors were separated to show tumor size. Frozen sections were prepared from tumor tissues. IF staining with antibodies to Ki-67 (in red color, (H)) and CC3 (in red color, (J)) was performed to detect proliferation and apoptosis of Panc-1 cells. And the percentage of Ki-67 + cells (I) or CC3 + cells (K) per field was shown by bar plots. * P < 0.05, * * P < 0.01, * ** P < 0.001. GEM, gemcitabine.

Article Snippet: Recombinant human complement C3a protein (MedChemExpress, MCE; HY-P7862), recombinant mouse complement C3a protein (MCE; HY-P7863) and C3aR antagonist, SB290157 (TargetMol) were used according to the manufacturer’s instructions.

Techniques: Activation Assay, Migration, CCK-8 Assay, Construct, Injection, Staining

Journal: Journal of Fungi

Article Title: Vaginal Clinical Isolates of Candida albicans Differentially Modulate Complosome Activation in Vaginal Epithelial Cells

doi: 10.3390/jof11070501

Figure Lengend Snippet: Intracellular C3aR and C5aR1 analysis. A monolayer of VECs was infected or not (Mock) with the Colonizing strain (Col) or VVC strain (VVC) for 4 h. After incubation, intracellular C3aR ( a ) and C5aR1 ( b ) were analyzed by cytofluorimetric analysis. Data in the graphs show the mean ± SEM of C3aR- and C5aR1-positive cells (cell counts from 8000 live cells) obtained from 4 different experiments. ns—not significant; * p ≤ 0.05.

Article Snippet: The anti-C3aR antibody was obtained from Miltenyi Biotech (Bergisch Gladbach, Germany).

Techniques: Infection, Incubation

Journal: Journal of Fungi

Article Title: Vaginal Clinical Isolates of Candida albicans Differentially Modulate Complosome Activation in Vaginal Epithelial Cells

doi: 10.3390/jof11070501

Figure Lengend Snippet: Activation of complosome in VECs by C. albicans . Differential activation of C3, C3a and C3b ( a ) and C5 and C5a ( b ) and intracellular C3aR and C5aR1 involvement in VECs infected with the Colonizing or VVC-associated strain. Arrows indicate changes in the levels of complement factors and receptors after infection with either the Colonizing strain (left) or the VVC strain (right). Green indicates increased levels and red indicates lower levels as compared to resting levels. Created with BioRender (adapted from King B.C. and Blom A.M., 2024 ).

Article Snippet: The anti-C3aR antibody was obtained from Miltenyi Biotech (Bergisch Gladbach, Germany).

Techniques: Activation Assay, Infection

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: As shown in Fig. c, d, the data from a single-cell transcriptome database indicated that the C3a receptor (C3aR) is strongly expressed in microglia (Tabula Muris).

Techniques: Double Immunostaining, Expressing, Cell Culture, Control