Figure 4

Immunofluorescence of a deparaffinized section of complete intestinal metaplasia of the stomach after immunostaining against Cnx (A, C, and E; green), lysozyme (B, C, and E; blue), and Ki67 (D and E; red); and serial sections of complete intestinal metaplasia after mRNA in situ hybridization (G to I) and DAB immunostaining against Cnx (H). A, Strong immunofluorescence of the Cnx protein (green) is found in a small number of Paneth cells identified by cosynthesis of lysozyme (blue; B) (arrowheads indicate Paneth cells). C, The overlay of Cnx (green) and lysozyme (blue) shows that in Paneth cells (arrowheads), a moderate immunostaining against Cnx is not regularly associated with a high density of lysozyme (arrow). D, Nuclear Ki67 protein (red) is found in several proliferating cells (arrowheads indicate Paneth cells). E, Triple immunofluorescence of lysozyme (blue), nuclear Ki67 (red), and Cnx (green) in a Paneth cell (arrow) of intestinal metaplasia. The majority of Ki67-producing cells (red) do not show any staining for lysozyme (blue), indicative for proliferating enterocytes (arrowheads indicate Paneth cells). F, Interference phase contrast view of the same section (arrowheads indicate Paneth cells). G, in situ hybridization for Cnx mRNA using digoxigenin-labeled antisense riboprobes. Cnx mRNA is found in Paneth cells (arrowheads). H, The same tissue section used for mRNA in situ hybridization (G) with DAB immunostaining against Cnx (arrowheads indicate Paneth cells). Immunostaining of Cnx is found in Paneth cells (arrowheads). I, Serial tissue section from G and H after mRNA in situ hybridization using digoxigenin-labeled sense riboprobes to Cnx (control; arrowheads indicate Paneth cells). Original magnification: A to F, ~ × 60; G to I, ~ × 60.