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To assess apoptosis, CD8+ T cells were isolated from splenocytes using a CD8α isolation kit (Miltenyi Biotec, Germany) according to the manufacturer's protocol, stimulated as described, stained with annexin V and PI (both BD Biosciences) according to the manufacture's protocol and then analysed by flow cytometry as described previously.

For a more detailed examination of the sperm heads, smears were prepared as described above, stained with carbolfuchsin-eosin according to the method described by [ 25] and modified by [ 15].

Primary cells from six patients were sorted as previously described and stained with an antibody against Ki67.

Sections were cut as described previously, stained with haematoxylin and eosin and examined to determine the exact location of the spheroid.

After 24 h cells were treated as described and stained with Annexin-V-PE (Bender MedSystem) according to the manufacturer's protocol.

Titanium discs were incubated with the bacterial co-culture as described above, stained with 10 μl LIVE/DEAD® (1% in sterile distilled water) (Molecular Probes, Stockholm, Sweden) and incubated in the dark for 20 minutes.

Blood cells were isolated on Ficoll as described and stained with CD14-FITC (Biolegend, San Diego, CA, USA) or MCSF-APC (Biolgened) or F4.80-FITC (Biolegend) antibodies at RT for 30 min.

As previously described, staining of human synovial tissue with haematoxylin and eosin showed an increase in the number of cell layers of the synovial lining of RA patients.

Mouse neutrophils were purified from bone marrow on Percoll gradients (52%/68%/78%) and cell survival in culture (in the presence of 10 ng/ml GM-CSF where indicated) was assayed essentially as previously described, using staining with Cell Tracker Green (CTG) (Invitrogen, Mulgrave, Australia) and PI (2  μg/ml).

Histological sections of muscle were prepared from wax-embedded material as described previously [6], stained with hematoxyline and eosin (H&E staining) and analyzed using light microscopy.

RPE/choroidal flatmounts injected with either AdCAGpA or AdCAGsCD59 and subjected to laser burn as described above were stained with an antibody against mouse C9.