Journal: Cell Death & Disease

Article Title: Trajectory modeling of endothelial-to-mesenchymal transition reveals galectin-3 as a mediator in pulmonary fibrosis

doi: 10.1038/s41419-021-03603-0

Figure Lengend Snippet: A Schematic diagram showing the functional exploration of C3ar1 and galectin-3 in IPF using lentivirus and galectin-3 interference (shRNA group), respectively. Alternatively, antagonists were used to block the binding affinity of C3a and galectin-3 (antagonist group). B , C Body weight and survival of IPF mice with galectin-3 shRNA lentivirus ( B ) (LV C3ar1 shRNA and LV galectin-3 shRNA, n = 5) or antagonists ( C ) (SB290157: antagonist of C3ar1; GB1107: antagonist of GB1107, n = 6). D , E HE and Masson’s trichrome staining of mouse lung sections following lentivirus or antagonist treatments. Scale bar = 500 μm (global view) or 50 μm (detailed view). F Semiquantitative morphological index/scoring of lung sections. The grade ranges from 1 (normal) to 8 (complete fibrosis). n = 30 for each group; * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The C3a inhibitor SB290157 (N(2)-{(2,2-diphenylethoxy) acetyl}-L-arginine; IC 50 = 200 nM; MedChemExpress, USA, 2.5 mg/ml) was prepared at 10% DMSO, 40% PEG300, 5% tween-80, and 45% saline.

Techniques: Functional Assay, shRNA, Blocking Assay, Binding Assay, 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 and B ) Immunofluorescence detection of C3 (red), Nestin (yellow), Nuclei (DAPI, blue), GFAP (green), or Olig2 (cyan) in the perivascular ( A ) or hypoxic ( B ) niches of murine RCAS-PDGFB– and RCAS-shp53–induced gliomas. ( C and 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 1 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 and H ) Pearson correlation coefficient between HALLMARK_COMPLEMENT and HALLMARK_HYPOXIA signatures in the TCGA GBMLGG ( G ) or TCGA GBM ( H ) data set. ( 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% O 2 ), hypoxia (1% O 2 ), or severe hypoxia (0.1% O 2 ). Data are shown as mean ± SEM. * P < 0.05, ** P < 0.01, or *** P < 0,001. Statistical tests were 1-way ANOVA, with Tukey post hoc test, or unpaired t test in case of 2 sample comparisons. Statistical tests were 1-way ANOVA ( I and K ), with Tukey post hoc test, or unpaired t test in case of 2-sample comparisons ( J ).

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

Techniques: Immunofluorescence, Expressing

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

Techniques: Expressing, RNA Sequencing, Comparison