Figure 6

Immunohistochemical analysis of markers of cell proliferation, cell lineage and cellular activation in spotted wolffish scleral skeleton lesion lesion-like tissue. IHCs staining (red) for PCNA (A), IgM (B), CD45 (C), phosphotyrosine (D) and Sox9 (E) were performed on sections adjacent to the safranin O-fast green stained section shown in Fig. 2I. Areas shown in (A), (C) and (E) encompass the region indicated by the yellow box in Fig. 2I, while areas shown in (B) and (D) encompass the region indicated by the white box in Fig. 2I. (A) Staining for PCNA appeared as a cellular nuclear staining in the perichondrium (p) and some areas consistent with the position of the maturing chondrocytes (ch) (see Supplemental Fig. 8B). (B) Staining for IgM appeared in both perichondrium (p) and in regions containing maturing chondrocytes (ch). (C) Staining for CD45 appeared in the perichondrium (p) and some areas consistent with the position of the maturing chondrocytes (ch) (see Supplemental Fig. 8A). (D) Staining for phosphotyrosine appeared in tracts in the perichondrium (p) projecting into the center of the lesion-like area and in areas consistent with the position of the maturing chondrocytes (ch) (see Supplemental Fig. 10). (E) Sox9 appeared as a cellular nuclear staining within the perichondrial (p) tracts projecting into the center of the lesion-like area, in areas consistent with the position of the maturing chondrocytes and in the cartilage (ca) (see supplemental Fig. 9). No counterstain was applied to the sections shown in (A–E) in order to emphasize the IHC signals. Scale grid subdivisions are 10 µm.