Fig 1.
Carboxy-terminal sequence of human, mouse, and rat C5aR1.
Differences in amino acid sequences of rat and mouse C5aR1 in comparison with the human receptor are marked in red.
Table 1.
Presence of C5aR1 in the tumour cells of different tumour entities.
Fig 2.
Analysis of the specificity of anti-human rabbit polyclonal antibody {5227}.
(A) Western blot analysis of whole cell preparations from stably C5aR1- or C5aR2-transfected HEK-293 cells. Receptors were enriched using wheat germ lectin agarose beads. Samples were separated on 7.5% SDS-polyacrylamide gels and blotted onto PVDF membranes. Membranes were then incubated with anti-human C5aR1 antibody {5227} or anti-human C5aR2 antibody {5236}, and blots were developed using enhanced chemiluminescence. Ordinate, migration of protein molecular weight markers (kDa). Note that {5227} and {5236} selectively detected only the respective targeted receptor and did not cross-react with the other receptor or other membrane proteins. (B) Immunocytochemistry of stably C5aR1- or C5aR2-transfected HEK-293 cells. Cells were fixed and immunofluorescently stained with anti-human C5aR1 antibody {5227} or anti-human C5aR2 antibody {5236}. Again, {5227} and {5236} selectively detected only the targeted receptor and did not cross-react with the other receptor. Note also, that prominent immunofluorescence for C5aR1 was localised at the plasma membrane. Representative results from one of three independent experiments are shown. Scale bar: 20 μm.
Fig 3.
Immunohistochemical detection of C5aR1 localisation in non-neoplastic human tissues (I).
Immunohistochemical staining (red-brown colour), counterstaining with haematoxylin. Scale bar: 100 μm (C, F), 50 μm (A, B, D, E, G-I). Arrows in (G): positive immune cells in the heart; arrows in (H): positive cells at the margin of the pancreatic islet; arrows in (I): positive Kupffer cells in the liver. (D-F) enlarged sections of (A-C).
Fig 4.
Immunohistochemical detection of C5aR1 localisation in non-neoplastic human tissues (II).
Immunohistochemical staining (red-brown colour), counterstaining with haematoxylin. Scale bar: 50 μm (A, D, C, E), 30 μm (D, F). Inset in (B): for adsorption control, the anti-C5aR1 antibody {5227} was incubated with 10 μg/ml of the peptide used for immunisations. (E and F) enlarged sections of (B and C).
Fig 5.
Immunohistochemical detection of C5aR1 localisation in human tumour entities (I).
Immunohistochemical staining (red-brown colour), counterstaining with haematoxylin. Scale bar: 50 μm (A-F). Insets in (A, C, D, E): for adsorption control, the anti-C5aR1 antibody {5227} was incubated with 10 μg/ml of the peptide used for immunisations. Lung SQCC: squamous cell carcinoma of the lung; PTC: papillary thyroid carcinoma; ATC: anaplastic thyroid carcinoma; CCC: cholangiocellular carcinoma.
Fig 6.
Immunohistochemical detection of C5aR1 localisation in human tumour entities (II).
Immunohistochemical staining (red-brown colour), counterstaining with haematoxylin. Scale bar: 50 μm (A-F). Insets in (A, B, D, E): for adsorption control, the anti-C5aR1 antibody {5227} was incubated with 10 μg/ml of the peptide used for immunisations. Arrow in (C): tumour cells (lymph node metastasis of a cervical carcinoma); arrowheads in (C): lymph follicle. MTS: lymph node metastasis; PSRC: pleomorphic sarcoma.