Journal: Oncotarget

Article Title: C5a receptor (CD88) promotes motility and invasiveness of gastric cancer by activating RhoA

doi: 10.18632/oncotarget.12656

Figure Lengend Snippet: A. , Western blots demonstrating the level of C5aR-expression in gastric cancer cell lines. B and C. , growth assays using Cell Counting Kit-8 showing the growth of C5aR positive and negative gastric cancer cells following stimulating with rC5a. C5aR: C5a receptor, PBMC: peripheral blood mononuclear cell, rC5a: recombinant C5a, GC: gastric cancer.

Article Snippet: A rabbit polyclonal antibody against C5aR (CD88; sc-25774) was purchased from Santa Cruz Biotechnology (Texas, USA).

Techniques: Western Blot, Expressing, Cell Counting, Recombinant

Journal: Oncotarget

Article Title: C5a receptor (CD88) promotes motility and invasiveness of gastric cancer by activating RhoA

doi: 10.18632/oncotarget.12656

Figure Lengend Snippet: A. , lower surfaces of an invasion membrane when MKN1 cells were assayed for invasion. B and C. , rC5a significantly enhanced the invasive ability of MKN1 and MKN7 cells. D. , rC5a did not enhance the invasive ability of AGS cells. E–H. , suppression of C5aR-expression using two kinds of siRNA significantly decreased the invasive ability of MKN1 and MKN7 cells. I and J. , W-54011, a C5aR-antagonist, significantly suppressed the invasive ability of MKN1 and MKN7 cells. C5aR, C5a receptor; rC5a, recombinant C5a; DMSO, Dimethyl sulfoxide; W-54011, C5aR-antagonist; *p<0.05; **p<0.01. NS, not significant.

Article Snippet: A rabbit polyclonal antibody against C5aR (CD88; sc-25774) was purchased from Santa Cruz Biotechnology (Texas, USA).

Techniques: Membrane, Expressing, Recombinant

Journal: Oncotarget

Article Title: C5a receptor (CD88) promotes motility and invasiveness of gastric cancer by activating RhoA

doi: 10.18632/oncotarget.12656

Figure Lengend Snippet: A. , Flow cytometry showed that C5aR proteins were overexpressed in the cellular membrane of NUGC3/C5aR cells. B. , rC5a significantly promotes the invasive ability of NUGC3/C5aR cells, but did not significantly promote the invasive ability of NUGC3/mock cells. C. , W-54011 significantly decreased the invasive ability of NUGC3/C5aR cells. D and E. , rC5a significantly promotes the mobility and the total distance of cell migration of NUGC3/C5aR cells but did not significantly promote the mobility of NUGC3/mock cells. F and G. , W-54011 significantly decreased the mobility of NUGC3/C5aR cells. C5aR, C5a receptor; rC5a, recombinant C5a; DMSO, Dimethyl sulfoxide; W-54011, C5aR-antagonist; **p<0.01. NS, not significant.

Article Snippet: A rabbit polyclonal antibody against C5aR (CD88; sc-25774) was purchased from Santa Cruz Biotechnology (Texas, USA).

Techniques: Flow Cytometry, Membrane, Migration, Recombinant

Journal: Oncotarget

Article Title: C5a receptor (CD88) promotes motility and invasiveness of gastric cancer by activating RhoA

doi: 10.18632/oncotarget.12656

Figure Lengend Snippet: A. , NUGC3/C5aR and NUGC3/mock cells were incubated with rC5a (10 nM) and fixed at the indicated times. F-actin was visualized by immunofluorescence staining with Alexa 488–conjugated phalloidin. Scale bars, 10 μm. Orange and yellow arrows and arrowheads indicate filopodia, stress fibers and membrane ruffling, respectively. B. , analysis of the activation of RhoA using a G-LISA on the lysates of NUGC3/C5aR and NUGC3/mock cells were extracted at the indicated times after rC5a treatment. C. , diagram of C5a-C5aR signaling via the RhoA pathway in gastric cancer cells. C5aR, C5a receptor; rC5a, recombinant C5a; GDP, guanosine diphosphate; GTP, guanosine triphosphate; *p<0.05; **p<0.01. NS, not significant.

Article Snippet: A rabbit polyclonal antibody against C5aR (CD88; sc-25774) was purchased from Santa Cruz Biotechnology (Texas, USA).

Techniques: Incubation, Immunofluorescence, Staining, Membrane, Activation Assay, Recombinant

Journal: Oncotarget

Article Title: C5a receptor (CD88) promotes motility and invasiveness of gastric cancer by activating RhoA

doi: 10.18632/oncotarget.12656

Figure Lengend Snippet: A. , gastric cancer tissues were immunohistochemically stained with anti-C5aR antibody. One-hundred cases were scored from 0 to 3 according to the extent of C5aR staining in the cancer area. Scale bar: 100μm. B. , Relapse-free survival curves for 86 patients who underwent a gastrectomy for gastric cancer (excluding stage IV patients), which were stratified by low- and high-expression of C5aR. C. , Overall survival curves for 100 patients who underwent gastrectomy for gastric cancer, which were stratified by low- and high-expression of C5aR. C5aR, C5a receptor; RFS, relapse-free survival; OS, overall survival.

Article Snippet: A rabbit polyclonal antibody against C5aR (CD88; sc-25774) was purchased from Santa Cruz Biotechnology (Texas, USA).

Techniques: Staining, Expressing

Journal: Oncotarget

Article Title: C5a receptor (CD88) promotes motility and invasiveness of gastric cancer by activating RhoA

doi: 10.18632/oncotarget.12656

Figure Lengend Snippet: Analysis of clinical factors associated with C5aR-expression from 100 patients with gastric cancer

Article Snippet: A rabbit polyclonal antibody against C5aR (CD88; sc-25774) was purchased from Santa Cruz Biotechnology (Texas, USA).

Techniques:

Journal: Oncotarget

Article Title: C5a receptor (CD88) promotes motility and invasiveness of gastric cancer by activating RhoA

doi: 10.18632/oncotarget.12656

Figure Lengend Snippet: Univariate and multivariate analysis of prognostic factors associated with the overall survival of patients with gastric cancer

Article Snippet: A rabbit polyclonal antibody against C5aR (CD88; sc-25774) was purchased from Santa Cruz Biotechnology (Texas, USA).

Techniques:

Journal: PLoS ONE

Article Title: Complement and the Alternative Pathway Play an Important Role in LPS/D-GalN-Induced Fulminant Hepatic Failure

doi: 10.1371/journal.pone.0026838

Figure Lengend Snippet: Wt C57BL/6 mice were injected i.p. with LPS/D-GalN and euthanized at 0, 1, 4 and 8 hours after the injection. (A–L) Immunohistochemical staining for C3, C3aR and C5aR in liver sections after the LPS/D-GalN injection. (M–N) The relative C3aR mRNA and C5aR mRNA expression levels were determined in live tissue at the indicated time points after the LPS/D-GalN injection. The mRNA expression was determined by relative quantitative real-time PCR analysis. The results are expressed as the means±SEM relative to GAPDH expression. (O) The serum concentrations of C3a at the indicated times after the LPS/D-GalN injection. ** and *** indicate p <0.01 and p <0.001, respectively. n = 6–7 per group. The original magnification for C3 of stained images: ×200; for C3aR and C5aR of stained images: ×800.

Article Snippet: The sections were incubated overnight at 4°C with rat anti-mouse C3mAb (1∶20 dilution, HyCult Biotechnology bv, Uden, Netherlands), rabbit anti-mouse C3aR polyclonal antibody(1∶50 dilution, Santa Cruz Biotechnology), rabbit anti-mouse C5aR polyclonal antibody (1∶80 dilution, Santa Cruz Biotechnology) and anti-C5b-9 polyclonal antibody (5 ug/ml, Calbiochem,SanDiego,CA).

Techniques: Injection, Immunohistochemical staining, Staining, Expressing, Real-time Polymerase Chain Reaction

Journal: PLoS ONE

Article Title: Complement and the Alternative Pathway Play an Important Role in LPS/D-GalN-Induced Fulminant Hepatic Failure

doi: 10.1371/journal.pone.0026838

Figure Lengend Snippet: (A–F) C3aR antagonist group mice displayed reduced liver damage (A–B, D–E) and decreased C3 deposition (C, F) 8 hours after LPS/D-GalN injection. (G–H) C3aR mRNA expression decreased at 8 hours compared with that of the saline group, whereas C5aR mRNA expression decreased from 4 to 8 hours (n = 4–5). (I) The different response patterns of ALT concentration in the C3aR antagonist group mice and the wt mice (n = 4–5). (J–L) The concentrations of TNF-α and IL-6 in the C3aR antagonist mice were lower than in the saline group. There was a delayed increase in MCP-1 in the C3aR antagonist group (n = 4–5). (M) Treatment with the C3aR antagonist increased the survival rate of the mice after LPS/D-GalN injection (n = 8). *, **and *** indicate p <0.05, p <0.01 and p <0.001, respectively, relative to the saline group. The means±SEM are shown. Magnification of the H&E and immunohistochemically stained images: ×200. The results are representative of 3 separate experiments.

Article Snippet: The sections were incubated overnight at 4°C with rat anti-mouse C3mAb (1∶20 dilution, HyCult Biotechnology bv, Uden, Netherlands), rabbit anti-mouse C3aR polyclonal antibody(1∶50 dilution, Santa Cruz Biotechnology), rabbit anti-mouse C5aR polyclonal antibody (1∶80 dilution, Santa Cruz Biotechnology) and anti-C5b-9 polyclonal antibody (5 ug/ml, Calbiochem,SanDiego,CA).

Techniques: Injection, Expressing, Saline, Concentration Assay, Staining

Journal: PLoS ONE

Article Title: Complement and the Alternative Pathway Play an Important Role in LPS/D-GalN-Induced Fulminant Hepatic Failure

doi: 10.1371/journal.pone.0026838

Figure Lengend Snippet: (A–I) Both C5aRmAb and CR2-fH groups displayed reduced liver damage (A–B, D–E, G–H) and decreased C3 deposition (C, F, I) 8 hours after LPS/D-GalN injection. (J) The different response patterns of ALT concentration in the C3aR antagonist mice and the wt mice (n = 4–5). (J–K) Treatment with the C5aR antagonist or CR2-fH increased the survival rate of the mice after LPS/D-GalN injection (n = 8). ** and *** indicate p <0.01 and p <0.001, respectively, relative to the saline group. The means±SEM are shown. Magnification of the H&E and immunohistochemically stained images: ×200. The results are representative of 3 separate experiments.

Article Snippet: The sections were incubated overnight at 4°C with rat anti-mouse C3mAb (1∶20 dilution, HyCult Biotechnology bv, Uden, Netherlands), rabbit anti-mouse C3aR polyclonal antibody(1∶50 dilution, Santa Cruz Biotechnology), rabbit anti-mouse C5aR polyclonal antibody (1∶80 dilution, Santa Cruz Biotechnology) and anti-C5b-9 polyclonal antibody (5 ug/ml, Calbiochem,SanDiego,CA).

Techniques: Injection, Concentration Assay, Saline, Staining