Journal: Redox Biology

Article Title: The C3–C3aR axis drives rotenone-induced cognitive damage via synaptic engulfment, dark microglia and PANoptosis

doi: 10.1016/j.redox.2026.104062

Figure Lengend Snippet: Inhibition of C3aR ameliorates rotenone-induced neurodegeneration, synaptic loss, α-synuclein phosphorylation and cognitive deficits in mice. C57BL/6 mice were treated with rotenone for 3 weeks to establish PD models, with 30 min of SB290157 (C3aR inhibitor, 1 mg/kg, i.p.) administration after each rotenone injection. (A) Concentrations of C3a in the hippocampus of Con and Rot mice, measured by enzyme-linked immunosorbent assay (ELISA). n = 6. (B) Representative Western blot images and densitometric quantification of C3aR protein levels in the hippocampus and rotenone mice. n = 4. (C) The mRNA levels of C3aR in vehicle and rotenone mice. (D) Representative images of double-immunofluorescence staining with C3aR and Iba-1 or Neu-N antibodies and (E, F) the quantification of C3aR + Iba-1 + and C3aR + Neu-N + cells in vehicle and rotenone mice. n = 3. (G) The quantification of Neu-N + cell number and (H) optical density of PSD95 staining in rotenone mice with or without C3aR inhibitor treatment. (I) The representative images of Neu-N and PSD95 staining in rotenone mice with or without C3aR inhibitor treatment. n = 6. (J, K) Representative bands of Neu-N, PSD95 and ser129-phosphorylated α-synuclein and the quantification of blots in rotenone mice with or without C3aR inhibitor treatment. n = 4. (L) Representative images of ser129-phosphorylated α-synuclein and (M) the quantification of staining density in rotenone mice with or without C3aR inhibitor treatment. n = 3. (N–Q) Escape latency, traveled distance, first platform crossing latency and platform crossing number in rotenone mice with or without C3aR inhibitor treatment. n = 15; ∗ p < 0.05, ∗∗ p < 0.01 for comparison between Rot & Rot + C3aR inhibitor groups; Scale bar = 50 μm.

Article Snippet: The C3a ELISA kit was purchased from CUSABIO (CSB- E08511 m, Houston, TX, USA), and TUNEL assay kit was provided by KeyGEN (KGA7073-1, Nanjing, China.

Techniques: Inhibition, Phospho-proteomics, Injection, Enzyme-linked Immunosorbent Assay, Western Blot, Double Immunofluorescence Staining, Staining, Comparison

Journal: Experimental and Therapeutic Medicine

Article Title: The complement C3a-C3aR and C5a-C5aR pathways promote viability and inflammation of human retinal pigment epithelium cells by targeting NF-κB signaling

doi: 10.3892/etm.2022.11420

Figure Lengend Snippet: Complement C3a and C5a aggravate inflammation in HRPE cells via the NF-κB signaling pathway. HRPE cells were treated with recombinant human complement component C3a (2 µg/ml) or C5a (1 µg/ml) with or without NF-κB inhibitor PDTC (10 µM), and the release of (A) TNF-α, (B) IL-1β, (C) IL-6, (D) PGE2 and (E) IL-10 was measured by ELISA. *** P<0.001 relative to control; ### P<0.001 relative to C3a or C5a treatment. PGE2, prostaglandin E2; HRPE, human retinal pigment epithelium.

Article Snippet: Following 24 h of treatment, the contents of various cytokines and other compounds were analyzed in HRPE cell supernatants and/or human vitreous humor were determined using the following ELISA kits in accordance with the manufacturers' protocols: Tumor Necrosis Factor-α Assay Kit (cat. no. H052-1), Interleukin-1β Assay Kit (cat. no. H002), Interleukin-6 Assay Kit (cat. no. H007-1-1) and Interleukin-10 Assay Kit (cat. no. H009-1; all from Nanjing Jiancheng Bioengineering Institute); prostaglandin E2 (PGE2) ELISA Kit (cat. no. E-EL-0034c; Elabscience Biotechnology Co., Ltd.); Human Complement Fragment 3a ELISA Kit (cat no. CSB-E08509h) and Human Complement Fragment 5a ELISA Kit (cat. no. CSB-E08512h; both from Cusabio).

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Control

Journal: Experimental and Therapeutic Medicine

Article Title: The complement C3a-C3aR and C5a-C5aR pathways promote viability and inflammation of human retinal pigment epithelium cells by targeting NF-κB signaling

doi: 10.3892/etm.2022.11420

Figure Lengend Snippet: Complement C3aR and C5aR antagonist inhibit inflammation and NF-κB signaling in HRPE cells challenged with complement C3a and C5a. HRPE cells were treated with recombinant human complement component C3a (2 µg/ml) and with or without C3aR antagonist SB290157 (20 µM), and the release of (A) TNF-α, (B) IL-1β, (C) IL-6, (D) PGE2 and (E) IL-10 was determined by ELISA. HRPE cells were treated with recombinant human complement component C5a (1 µg/ml) and with or without C5aR antagonist CCX168 (2 µM), and the release of (F) TNF-α, (G) IL-1β, (H) IL-6, (I) PGE2 and (J) IL-10 was determined by ELISA. (K) The phosphorylation of NF-κB and expression of NF-κB in HRPE cells treated with recombinant human complement component C3a (2 µg/ml) and with or without C3aR antagonist SB290157 (20 µM) were determined by western blot. (L) The phosphorylation of NF-κB and expression of NF-κB in HRPE cells treated with recombinant human complement component C5a (1 µg/ml) and with or without C5aR antagonist CCX168 (2 µM) were determined by western blot. *** P<0.001 relative to control; ### P<0.001 relative to C3a or C5a treatment. p-NF-κB, phosphorylated NF-κB; C5aR, C5a receptor; HRPE, human retinal pigment epithelium; CCX, CCX168; SB, SB290157.

Article Snippet: Following 24 h of treatment, the contents of various cytokines and other compounds were analyzed in HRPE cell supernatants and/or human vitreous humor were determined using the following ELISA kits in accordance with the manufacturers' protocols: Tumor Necrosis Factor-α Assay Kit (cat. no. H052-1), Interleukin-1β Assay Kit (cat. no. H002), Interleukin-6 Assay Kit (cat. no. H007-1-1) and Interleukin-10 Assay Kit (cat. no. H009-1; all from Nanjing Jiancheng Bioengineering Institute); prostaglandin E2 (PGE2) ELISA Kit (cat. no. E-EL-0034c; Elabscience Biotechnology Co., Ltd.); Human Complement Fragment 3a ELISA Kit (cat no. CSB-E08509h) and Human Complement Fragment 5a ELISA Kit (cat. no. CSB-E08512h; both from Cusabio).

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Phospho-proteomics, Expressing, Western Blot, Control

Journal: JCI Insight

Article Title: C3a and suPAR drive versican V1 expression in tubular cells of focal segmental glomerulosclerosis

doi: 10.1172/jci.insight.122912

Figure Lengend Snippet: (A) RT-PCR analysis of total versican in tubular cells treated with 20% patient serum (PS), 20% heat-inactivated patient serum (HIPS), 20% healthy serum (HS), or 20% heat-inactivated healthy serum (HIHS) for 48 hours (n = 3). (B) RT-PCR analysis of total versican in tubular cells treated with 20% PS, eculizumab, or SB290157 for 48 hours (n = 5). (C) Level of serum C3a in FSGS patients and controls (n = 20). (D) Level of urinary C3a in FSGS patients and controls (n = 20). (E) Immunofluorescence staining of C3a (red) and C3aR (green) in renal tissue of FSGS patients (n = 3). (F) RT-PCR analysis of total versican, versican V1, V0, and V3 in tubular cells treated with C3a or 20% PS for 48 hours (n = 3). Scale bars: 20 μm. For statistical analysis, 1-way ANOVA with Tukey’s post hoc test was used for A, B, and F, and a 2-tailed Student’s t test was used for C and D. *P < 0.05 compared with control; #P < 0.05 compared with PS-, HS-, or C3a-treated cells.

Article Snippet: A sandwich ELISA was performed according to the manufacturer’s protocol to quantify human C3a (KA1020, Novus Biologicals), mouse C3a (NBP2-70037, Novus Biologicals), human uPAR (DY807, R&D Systems), and mouse uPAR (DY531, R&D Systems).

Techniques: Reverse Transcription Polymerase Chain Reaction, Immunofluorescence, Staining, Control

Journal: JCI Insight

Article Title: C3a and suPAR drive versican V1 expression in tubular cells of focal segmental glomerulosclerosis

doi: 10.1172/jci.insight.122912

Figure Lengend Snippet: (A) RT-PCR analysis of total versican in tubular cells treated with C3a and SB203580, PD098059, or MK2206 (n = 5). (B) RT-PCR analysis of versican V1, V0, and V3 in tubular cells treated with C3a and MK2206 (n = 3). (C) Western blot analysis of phospho-AKT (p-AKT) in tubular cells (n = 3). (D) Western blot analysis of nuclear β-catenin in tubular cells (n = 3). (E) Schematic of the β-catenin/TCF binding sites in the upstream sequence of the versican promoter and the constructed versican promoter–luciferase reporter plasmids. (F) ChIP analysis of the binding between β-catenin and the versican promoter in tubular cells treated with C3a (n = 3). (G) Normalized luciferase activity of reporter constructs in tubular cells cotransfected with N90-β-catenin plasmid (n = 3). (H) RT-PCR analysis of total versican, versican V1, V0, and V3 in tubular cells transfected with N90-β-catenin plasmid (n = 3). (I) RT-PCR analysis of total versican, versican V1, V0, and V3 in tubular cells treated with C3a and si-CTNNB1 (n = 3). For statistical analysis, 1-way ANOVA with Tukey’s post hoc test was used for A, B, and I, and a 2-tailed Student’s t test was used for G and H. *P < 0.05 compared with control; #P < 0.05 compared with C3a-treated cells.

Article Snippet: A sandwich ELISA was performed according to the manufacturer’s protocol to quantify human C3a (KA1020, Novus Biologicals), mouse C3a (NBP2-70037, Novus Biologicals), human uPAR (DY807, R&D Systems), and mouse uPAR (DY531, R&D Systems).

Techniques: Reverse Transcription Polymerase Chain Reaction, Western Blot, Binding Assay, Sequencing, Construct, Luciferase, Activity Assay, Plasmid Preparation, Transfection, Control

Journal: JCI Insight

Article Title: C3a and suPAR drive versican V1 expression in tubular cells of focal segmental glomerulosclerosis

doi: 10.1172/jci.insight.122912

Figure Lengend Snippet: (A) Level of serum creatinine in WT and C3aR-knockout (C3aR–/–) mice treated with Adriamycin (ADR) (n = 6). (B and C) Masson’s trichrome staining of renal sections in WT and C3aR–/– mice treated with ADR (n = 6). (D) RT-PCR analysis of total versican, versican V1, V0, and V3 in tubulointerstitial tissues of WT and C3aR–/– mice treated with ADR (n = 6). (E and F) Immunohistochemical staining of versican in renal tissues of WT and C3aR–/– mice treated with ADR (n = 6). (G) Level of urinary C3a in WT and C3aR–/– mice treated with ADR (n = 6). (H) Immunofluorescence staining of C3a in WT and C3aR–/– mice treated with ADR (n = 6). (I) Western blot analysis of p-AKT in tubulointerstitial tissues of WT and C3aR–/– mice treated with ADR (n = 6). (J) Western blot analysis of nuclear β-catenin (n = 6). (K) ChIP analysis of the binding between β-catenin and the versican promoter in tubulointerstitial tissues (n = 6). (L) Western blot analysis of versican V1 in tubulointerstitial tissues (n = 6). Scale bars: 20 μm (E and H). Scale bars: 30 μm (B). For statistical analysis, 1-way ANOVA with Tukey’s post hoc test was used for A, C, D, F, and G. *P < 0.05 compared with control WT mice; #P < 0.05 compared with ADR-treated WT mice.

Article Snippet: A sandwich ELISA was performed according to the manufacturer’s protocol to quantify human C3a (KA1020, Novus Biologicals), mouse C3a (NBP2-70037, Novus Biologicals), human uPAR (DY807, R&D Systems), and mouse uPAR (DY531, R&D Systems).

Techniques: Knock-Out, Staining, Reverse Transcription Polymerase Chain Reaction, Immunohistochemical staining, Immunofluorescence, Western Blot, Binding Assay, Control

Journal: JCI Insight

Article Title: C3a and suPAR drive versican V1 expression in tubular cells of focal segmental glomerulosclerosis

doi: 10.1172/jci.insight.122912

Figure Lengend Snippet: (A) Level of serum suPAR in FSGS patients (n = 20). (B) Level of urinary suPAR in FSGS patients (n = 20). (C) Immunohistochemical staining of uPAR in renal tissues of FSGS patients (n = 5). (D) RT-PCR analysis of total versican, versican V1, V0, and V3 in tubular cells treated with 20% PS and uPAR-blocking antibody (n = 5). (E) RT-PCR analysis of total versican, versican V1, V0, and V3 in tubular cells treated with C3a and suPAR (n = 5). (F) RT-PCR analysis of ITGB6 in tubulointerstitial tissues of FSGS patients (n = 20). (G) IP analysis of the binding between suPAR and ITGB6 in tubular cells treated with 20% PS (n = 3). (H) Immunofluorescence staining of uPAR (green) and ITGB6 (red) in tubular cells treated with 20% PS (n = 3). Scale bars: 20 μm (C and H). (I) RT-PCR analysis of total versican, versican V1, V0, and V3 in tubular cells treated with 20% PS and si-ITGB6 (n = 5). (J) Rac1 activation assay in tubular cells treated with 20% PS and si-ITGB6 (n = 3). (K) RT-PCR analysis of total versican, versican V1, V0, and V3 in tubular cells treated with 20% PS and si-Rac1 (n = 5). For statistical analysis, a 2-tailed Student’s t test was used for A, B, and F, and 1-way ANOVA with Tukey’s post hoc test was used for D, E, I, J, and K. *P < 0.05 compared with control; #P < 0.05 compared with PS- or C3a-treated cells.

Article Snippet: A sandwich ELISA was performed according to the manufacturer’s protocol to quantify human C3a (KA1020, Novus Biologicals), mouse C3a (NBP2-70037, Novus Biologicals), human uPAR (DY807, R&D Systems), and mouse uPAR (DY531, R&D Systems).

Techniques: Immunohistochemical staining, Staining, Reverse Transcription Polymerase Chain Reaction, Blocking Assay, Binding Assay, Immunofluorescence, Activation Assay, Control

Journal: JCI Insight

Article Title: C3a and suPAR drive versican V1 expression in tubular cells of focal segmental glomerulosclerosis

doi: 10.1172/jci.insight.122912

Figure Lengend Snippet: (A) Splicing factors predicted to bind to the 5′ end of versican exon 7 with the SpliceAid 2 database. (B) IP analysis of the binding between Rac1 and SRp40 in tubular cells treated with 20% PS (n = 3). (C) Immunofluorescence staining of SRp40 (green), Rac1 (red), and DAPI (blue) in tubular cells treated with 20% PS (n = 3). Scale bars: 20 μm. (D) RIP analysis of the binding of SRp40 and Rac1 to the 5′ end of versican exon 7 in tubular cells treated with 20% PS (n = 5). (E) RIP analysis of the binding of Rac1 to the 5′ end of versican exon 7 in tubular cells treated with 20% PS and si-SRp40 (n = 3). (F) RIP analysis of the binding of SRp40 to the 5′ end of versican exon 7 in tubular cells treated with 20% PS and si-Rac1 (n = 3). (G) Rac1 activity in tubular cells treated with 20% PS and si-SRp40 (n = 3). (H) RIP analysis of the binding of U2AF1 to the 3′ splice site of versican intron 6 in tubular cells treated with 20% PS and pGEMT-SRp40 plasmid or suPAR-blocking antibody (n = 5). (I) PCR analysis of versican V0 and V1 in tubular cells treated with 20% PS and pGEMT-SRp40 plasmid or suPAR-blocking antibody (n = 3). (J) RIP analysis of the binding of U2AF1 to the 3′ splice site of versican intron 6 in tubular cells treated with C3a and Rac1Q61L plasmid or suPAR (n = 5). (K) PCR analysis of versican V0 and V1 in tubular cells treated with C3a and Rac1Q61L plasmid or suPAR (n = 3). For statistical analysis, 1-way ANOVA with Tukey’s post hoc test was used for D–F, G, H, and J. *P < 0.05 compared with control; #P < 0.05 compared with PS- or C3a-treated cells.

Article Snippet: A sandwich ELISA was performed according to the manufacturer’s protocol to quantify human C3a (KA1020, Novus Biologicals), mouse C3a (NBP2-70037, Novus Biologicals), human uPAR (DY807, R&D Systems), and mouse uPAR (DY531, R&D Systems).

Techniques: Binding Assay, Immunofluorescence, Staining, Activity Assay, Plasmid Preparation, Blocking Assay, Control

Journal: JCI Insight

Article Title: C3a and suPAR drive versican V1 expression in tubular cells of focal segmental glomerulosclerosis

doi: 10.1172/jci.insight.122912

Figure Lengend Snippet: (A) Model of the 34-nt complementary sequence in intron 6 (site 1) and intron 8 (site 2) of versican pre-mRNA. (B) RAP analysis of the intron 6/8 interaction in versican pre-mRNA of tubular cells (n = 3). (C) RAP analysis of the intron 6/8 interaction in versican pre-mRNA of tubular cells treated with LNA antisense oligonucleotides targeting either site 1 or site 2 (n = 3). (D) PCR analysis of versican V3 in tubular cells (n = 3). (E) RAP analysis of the intron 6/8 interaction in versican pre-mRNA of tubular cells transfected with Rac1Q61L plasmid and si-SRp40 (n = 3). (F) PCR analysis of versican V3 in tubular cells transfected with Rac1Q61L plasmid and si-SRp40 (n = 3). (G) RAP analysis of the intron 6/8 interaction in versican pre-mRNA of tubular cells treated with 20% PS and si-Rac1 or suPAR-blocking antibody (n = 3). (H) PCR analysis of versican V3 in tubular cells treated with 20% PS and si-Rac1 or suPAR-blocking antibody (n = 3). (I) RAP analysis of the intron 6/8 interaction in versican pre-mRNA of tubular cells treated with C3a and Rac1Q61L plasmid or suPAR (n = 3). (J) PCR analysis of versican V3 in tubular cells treated with C3a and Rac1Q61L plasmid or suPAR (n = 3).

Article Snippet: A sandwich ELISA was performed according to the manufacturer’s protocol to quantify human C3a (KA1020, Novus Biologicals), mouse C3a (NBP2-70037, Novus Biologicals), human uPAR (DY807, R&D Systems), and mouse uPAR (DY531, R&D Systems).

Techniques: Sequencing, Transfection, Plasmid Preparation, Blocking Assay

Journal: JCI Insight

Article Title: C3a and suPAR drive versican V1 expression in tubular cells of focal segmental glomerulosclerosis

doi: 10.1172/jci.insight.122912

Figure Lengend Snippet: C3a promotes the transcription of versican by activating the AKT/β-catenin pathway in tubular cells. suPAR binds to ITGFB6 and activates Rac1, which translocates into the nucleus and binds to SRp40 at the 5′ end of exon 7 of versican pre-mRNA. This binding not only inhibits the 3′-end splicing of intron 6 but also inhibits the base-pair interaction between intron 6 and intron 8, which leads to the formation of versican V1.

Article Snippet: A sandwich ELISA was performed according to the manufacturer’s protocol to quantify human C3a (KA1020, Novus Biologicals), mouse C3a (NBP2-70037, Novus Biologicals), human uPAR (DY807, R&D Systems), and mouse uPAR (DY531, R&D Systems).

Techniques: Binding Assay

Journal: bioRxiv

Article Title: Engineering sialylation in pigs: A promising strategy for overcoming xenograft rejection and revolutionizing organ transplantation

doi: 10.1101/2025.04.30.651299

Figure Lengend Snippet: Schematic illustration of the in-vitro experimental procedure, measurements of human thrombin/anti-thrombin III complex (TAT), C3a, C4b, and C5b9 were taken from plasma in mixed human blood and porcine kidney and aorta tissues transfected with ST8Sia6. These were compared to controls porcine kidney and aorta endothelium transfected with GFP under in vitro conditions at three different time points (0, 3, and 6 hours). Result indicates that control GFP transfected Kidney and Aorta group (grey line) had increased complement activation. b-c, indicate C3a level in kidney and aorta condition respectively, similarly d-e, represent C4b level, f-g, represent C5b9 level from kidney and aorta experimental conditions. h-i, TAT level in plasma from kidney and aorta experimental conditions. In contrast, the ST8Sia6-transfected porcine kidney group (red line) showed a slight reduction in complement activation. Notably, the ST8Sia6- transfected aorta cells demonstrated significant reductions in complement activations. All graphs summarize the results from 4 donors (mean ± S.E) with 4 individual replicates. Significance differences in P value (P<0.05) were evaluated using ANOVA, and the P values are represented in the graphs.

Article Snippet: Plasma levels of thrombin/anti-thrombin III complex (TAT), C3a, C4d, and sC5b-9 were measured using commercially available ELISA kits: Human Thrombin-Antithrombin Complex ELISA Kit (Abcam AB1089071), Human C4b (Novus Biologicals™ NBP270046), Human C3a (Novus Biologicals™ NBP266755), and Human C5b-9 (Novus Biologicals™ NBP266708).

Techniques: In Vitro, Clinical Proteomics, Transfection, Control, Activation Assay

Journal: Results in Pharma Sciences

Article Title: Safety of poly (ethylene glycol)-coated perfluorodecalin-filled poly (lactide-co-glycolide) microcapsules following intravenous administration of high amounts in rats

doi: 10.1016/j.rinphs.2014.04.001

Figure Lengend Snippet: Effect of capsule infusion on release of cytokines and complement factors. PFD-filled PLGA microcapsules (1.5 µm), 0.25% PVA or 0.9% NaCl were infused for 30 min (20 ml/kg body weight × h). The values plotted are mean ± SEM of 6-4 (capsules) or 3 (PVA and NaCl) individual experiments, *p < 0.05 compared to NaCl group. The detection limit of the used ELISA was 0.08 ng/ml. Note: logarithmic scale was used for Fig. 4E, F and H. C4a level in NaCl and PVA groups at all time-points measured were below detection limit.

Article Snippet: The plasma concentration of complement factors 3 (C3) and 4a (C4a) were assessed with rat-specific ELISA kits (Rat complement fragment 4a ELISA kit, Cusabio, Wuhan, China, and Rat Complement Factor 3, GenWay Biotech Inc, San Diego, CA, USA) according to the manufacturer's instructions.

Techniques: Capsules, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: Zwitterionic hydrogel designs for conducting polymers enable bioelectronics with suppressed foreign body responses

doi: 10.1101/2025.10.31.683260

Figure Lengend Snippet: a , Total counts of macrophages associated with each type of implant, determined by flow cytometry and 4-week post-implantation. Macrophages were defined as F4/80 + cells. ZIPH-3c is consistently associated with the greatest number of macrophages. b , iNOS + Arg-1 ‒ / iNOS ‒ Arg-1 + macrophage ratios associated with each implant, determined by flow cytometry at 4 weeks post-implantation. iNOS + Arg-1 ‒ macrophages are considered to be M1-like and iNOS ‒ Arg-1 + macrophages are considered to be M2-like. ZIPH-3c is associated with the most M1-like macrophages. c , CD86 + CD206 ‒ / CD86 ‒ CD206 + macrophage ratios associated with each implant, determined by flow cytometry at 4 weeks post-implantation. CD86 + CD206 ‒ macrophages are considered to be M1-like and CD86 ‒ CD206 + macrophages are considered to be M2-like. ZIPH-3c is associated with the most M1-like macrophages. d , Representative immunofluorescence microscopy images of tissues stained with F4/80 (macrophage), CD86 (M1), and CD206 (M2). Locations of implants appear clear and are marked by asterisks. e , Percentage of CD3 + T cells that are T-bet + . T cells associated with ZIPH-3c have a high fraction of T-bet + cells. f , Percentage of CD4 + T helper cells that are FoxP3 + T reg s. CD4+ T helper cells associated with ZIPH-3c have a high fraction of FoxP3 + T reg s. g-h , Key cytokines associated with each implant, determined via LegendPlex assay at 1-week (g-h) and 4-week (i-j) post-implantation. MCP-1 is consistently upregulated for both ZIPH-3c and ZIPH-u. k , Total counts of neutrophils associated with each implant, determined by flow cytometry 1-week post-implantation. Neutrophils were defined as Ly-6G + cells. l , Complement C3a concentration associated with each implant, determined by ELISA 1-week post-implantation. The high number of neutrophils is correlated with the generation of C3a. PSB has a distinctly higher neutrophil number and C3a concentration than the rest of the hydrogels that contain PSS. Error bars, mean ± s.d. N = 5-6 mice per treatment. One-way ANOVA was used for statistical comparison of multiple means. *, P < 0.05; **, P < 0.01; ****, P < 0.0001; ##, P < 0.01 compared to rest; ####, P < 0.0001 compared to rest.

Article Snippet: A C3a ELISA kit (Novus Biologicals), a decorin ELISA kit (R & D Systems), and an endostatin ELISA kit (Abcam) were used following manufacturer’s instructions.

Techniques: Flow Cytometry, Immunofluorescence, Microscopy, Staining, Concentration Assay, Enzyme-linked Immunosorbent Assay, Comparison