recombinant mouse c3a  (R&D Systems)

Journal: Cells

Article Title: House Dust Mite Nebulization Drives Alarmin and Complement Activation in a Murine Tracheal Air–Liquid Interface Culture System

doi: 10.3390/cells14201598

Figure Lengend Snippet: Time-dependent complement activation and anaphylatoxin receptor expression in ALI cultures exposed to HDM. ( a ) C3 and C3a concentrations in cell lysates and the SN of unstimulated (steady-state) ALI cultures and 24, 48 and 72 h after HDM exposure (steady-state, 24, 48 and 72 h: n = 6). * p < 0.05; ** p < 0.01; *** p < 0.001. ( b ) HDM and HI-HDM mediated cleavage of hC3 into hC3a ( n = 3 independent experiments). Data are shown as mean ± SEM. Data were analyzed using an unpaired T -test. *** p < 0.001. ( c ) C5 and C5a concentrations in cell lysates and the SN of steady-state ALI cultures and 24, 48 and 72 h after HDM exposure. Data are shown as mean ± SEM (steady-state, 24, 48 and 72 h: n = 6). ( d ) Comparison of C3 and C5 ( left panel ), C3a and C5a ( right panel ) concentrations in cell lysates from steady-state ALI cultures and 24, 48 and 72 h after HDM exposure. Data are shown as mean ± SEM (steady-state, 24, 48 and 72 h: n = 6). Data were analyzed using two-way ANOVA with Šidák’s multiple comparisons test. **** p < 0.0001. ( e ) Immunofluorescence analysis of C3aR and C5aR1 expression in unstimulated ALI cultures and 24, 48 and 72 h after HDM exposure. Data are shown as mean ± SEM ( n = 3). Data were analyzed using one-way ANOVA with Holm–Šidák’s multiple comparisons test. * p < 0.05; ** p < 0.01. The number of experiments shown in ( a , c – e ) refers to biological replicates derived from 6 independent preparations of trachea, each representing a separate cell isolation and culture.

Article Snippet: Plates were washed four times with 0.05% Tween 20 in PBS; serial dilutions of recombinant mouse C3a (R&D Systems #8085-C3-025—Minneapolis, MN, USA) were performed; and samples were added (25 μL/well) and incubated at RT for 90 min. After washing four times with 0.05% Tween 20 in PBS, 25 μL/well of 1 μg/mL in 1% BSA/PBS biotinylated anti-mouse C3a detection antibody (clone I87-419, BD Biosciences #558251—San Jose, CA, USA) was added and incubated for 1 h at RT.

Techniques: Activation Assay, Expressing, Comparison, Immunofluorescence, Derivative Assay, Cell Isolation

Journal: Journal of the American Heart Association

Article Title: Hepatic Abnormal Secretion of Apolipoprotein C3 Promotes Inflammation in Aortic Dissection

doi: 10.1161/jaha.124.037172

Figure Lengend Snippet: Figure 1. Apo C3 is abundant in the plasma of patients with AD and deposits in the aorta. A, Heat map results of plasma exosome sequencing in patients with AD (n=20) and normal controls (n=20). B, ELISA results of apo C3 in the plasma of patients with AD (n=20) and NCs (n=20). C, Representative western blot of apo C3 protein in the aortas of patients with AD (n=6) and NCs (n=6). D, Representative immunohistochemistry of apo C3 in the aortas of patients with AD (n=8) and normal controls (n=8). E, Representative immunofluorescence of apo C3 in the aortas of patients with AD (n=9) and normal controls (n=9). F through H, Quantitative protein levels of apo C3 in western blot (C), immunohistochemistry (D), and immunofluorescence (E). Data are expressed as mean±SEM. Bars represent the means, and caps represent the SEM. Statistical comparisons were made using an unpaired t test (B, F through H). A value of P<0.05 was considered significant. The layers of the aorta are denoted as I for intima, M for media, and A for adventitia. AD indicates aortic dissection; apo C3 apolipoprotein C3; α-SMA, α-smooth muscle actin; CD: cluster of differentiation; ENO, enolase; LPL, lipoprotein lipase; MMP, matrix metalloproteinase; NC, normal control; and RBP, retinol-binding protein.

Article Snippet: Recombinant Mouse Apolipoprotein C3 (CUSABIO; CSB- EP001933MO) was added to MOVAS, MAECs, and J774A.1 cells at different concentrations and incubated for 48 hours.

Techniques: Clinical Proteomics, Sequencing, Enzyme-linked Immunosorbent Assay, Western Blot, Immunohistochemistry, Immunofluorescence, Dissection, Control, Binding Assay

Journal: Journal of the American Heart Association

Article Title: Hepatic Abnormal Secretion of Apolipoprotein C3 Promotes Inflammation in Aortic Dissection

doi: 10.1161/jaha.124.037172

Figure Lengend Snippet: Figure 3. Transcriptome sequencing reveals inhibition of TLR2/NLRP3 pathway activation, M1 macrophage polarization, and MMP release in the aortas of mice after hepatic apo C3 interference. A through E, Results of transcriptome sequencing in the aortas of mice in the BAPN group (n=5) and BAPN+Sh-apo C3 group (n=5). A, Volcano plot. B, Heat map. C, Protein–protein interaction network of apo C3 with differentially expressed genes. D, GO analysis results. E, KEGG analysis results. apo C3 indicates apolipoprotein C3; BAPN, β-aminopropionitrile; GO, gene ontology; KEGG, Kyoto Encyclopedia of Genes and Genomes; MMP, matrix metalloproteinase; NLRP3, NOD-like receptor pyrin domain containing 3; and TLR2, Toll-like receptor 2.

Article Snippet: Recombinant Mouse Apolipoprotein C3 (CUSABIO; CSB- EP001933MO) was added to MOVAS, MAECs, and J774A.1 cells at different concentrations and incubated for 48 hours.

Techniques: Sequencing, Inhibition, Activation Assay