Fig 4. Cytochemistry of cultured long bone marrow cells from EBV-infected and -uninfected hu-NOG mice in the presence of M-CSF and RANKL differentiation signals.
(A–D) Cultured cells from an EBV-infected mouse (NOG 12–6). (A) Cultured cells stained for TRAP. TRAP in the cultured multinucleated cell stained red violet. (B) Cultured cells immunostained for human cathepsin K. The positive multinucleated cell stained brown with the anti-human cathepsin K antibody (dilution, 1:1800). (C) Cultured cells immunostained for human mitochondrial protein. Multinucleated cell positively stained brown with anti-human mitochondria antibody (dilution, 1:40). (D) Cultured cells immunostained using isotype control rabbit immunoglobulin (dilution, 1:4500). (E–H) Cultured cells from another EBV-infected mouse (NOG 12–7). The multinucleated cells positively stained for TRAP (E), and with anti-human cathepsin K antibody (dilution, 1:600) (F) and anti-human mitochondria antibody (dilution, 1:10) (G). (H) Cultured cells immunostained using isotype control mouse immunoglobulin (dilution, 1:5). (I) Cultured cells from another EBV-infected mouse (NOG 18–1) on a pit formation assay plate. TRAP in the cultured multinucleated cells stained red violet. Calcium phosphate coating around the TRAP-positive multinucleated cell was eliminated (arrow). (J and K) Cultured cells from an EBV-uninfected mouse (NOG 24–1). (J) Cultured cells immunostained for human MMP-9. The multinucleated cell positive for human MMP-9 stained brown with anti-human MMP-9 antibody (dilution, 1:25). (K) Cultured cells immunostained for human mitochondrial protein. The multinucleated cell positive for human mitochondrial protein stained brown with anti-human mitochondria antibody (dilution, 1:10). (L and M) Cultured cells from another EBV-uninfected mouse (NOG 26–1). The multinucleated cells were considered positive when stained with anti-human MMP-9 antibody (dilution, 1:50) (L) and anti-human mitochondria antibody (dilution, 1:40) (M). (original magnification, 400×) Republished from [Nagasawa Y, Ikumi N, Nozaki T, Inomata H, Imadome K, et al. Human osteoclasts are mobilized in erosive arthritis of Epstein-Barr virus-infected humanized NOD/Shi-scid/IL2Rγnull mice. 2014 ACR/ARHP Annual Meeting. Abstract number:2340.] under a CC BY license, with permission from [American College of Rheumatology], original copyright [2014].