Journal: The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society

Article Title: Increased immunoreactivity of cathepsins in the rat esophagus under chronic acid reflux esophagitis.

doi: 10.1369/0022155414542300

Figure Lengend Snippet: Figure 2. Distribution of cathepsin B (CB) in the esophageal mucosa of sham (A–D) and Reflux Esophagitis (RE) (E–G) groups and the fundus ulcer of the RE group (H–J) at 28 days post-operatively. Triple staining for CB (green), lamp1 (red) and CD68 (blue) in addition to counterstaining with DAPI (white). Boxed areas in C and G are enlarged and shown as insets in C, and H and I, respectively. In the sham group, granular immunoreactivity for CB is localized intensely in the granular layer (g), only weakly localized in the basal layer (b), and absent in the spinous layer (s) (A–C). In the RE group, intense granular staining for CB is observed in the basal and spinous layers in addition to the granular layer (E–G). In both groups, CD68-positive macrophages are intensely positive for CB (arrowheads in A–D and H–J). Dotted lines: upper and lower limits of granular and basal layers, respectively. c: cornified layer, lp: lamina propria. Scale bars: 20 µm (A–G, J–L), 50 µm (inset in C, H, I). (M) Immunoblot analysis of CB in esophageal mucosae of the sham (S) and RE (R) groups. In both groups, the proform (pro) of CB (around 40 kD) is only weakly detected, whereas mature forms of CB (29 kD and 26 kD for single- and heavy-chain forms, respectively) are evident. Protein amount of both the proform and mature forms for CB is increased in the RE group.

Article Snippet: For the triple staining of CB, CD, CH or CL with lysosomal-associated membrane protein 1 (lamp1) and CD68, the sections were incubated with rabbit anti CB (06-480; Millipore, Temecula, CA, USA) (1:100), CD (Koike et al. 2005; Koike et al. 2013a) (1:100), CH (Kominami et al. 1985) (1:100), or CL (Bando et al. 1986), along with goat anti-lamp1 (AF4800, R&D Systems, Minneapolis, MN) (1:100), and mouse anti-CD68 (1:100).

Techniques: Reflux, Staining, Western Blot

Journal: The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society

Article Title: Increased immunoreactivity of cathepsins in the rat esophagus under chronic acid reflux esophagitis.

doi: 10.1369/0022155414542300

Figure Lengend Snippet: Figure 3. Distribution of cathepsin C (CC) in the esophageal mucosa of sham (A–D) and Reflux Esophagitis (RE) (E–G) groups and the fundus ulcer of the RE group (H–J) at 28 days post-operatively. Triple staining for CC (green), lamp1 (red) and CD68 (blue) in addition to counterstaining with DAPI (white). Boxed areas in C and G are enlarged and shown as insets in C, and H and I, respectively. In the sham group, granular immunoreactivity for CC is localized intensely in the granular layer (g), and moderately localized in the basal (b) and spinous (s) layers (A–C). In the RE group, intense granular staining for CC is observed in the basal and spinous layers in addition to the granular layer (E–G). In the sham group, some CD68-positive macrophages are intensely positive for CC (arrowheads in A–D), whereas some are negative for CC (arrows in A–D). In the RE group, most of the CD68-positive macrophages are intensely positive for CC (H–J). Dotted lines: upper and lower limits of granular and basal layers, respectively. c: cornified layer, lp: lamina propria. Scale bars: 20 µm (A–G, J–L), 50 µm (inset in C, H, I). (M) Immunoblot analysis of CC in esophageal mucosae of the sham (S) and RE (R) groups. The mature form of CC (25 kD for the heavy-chain form) is increased significantly in the mucosa of the RE group.

Article Snippet: For the triple staining of CB, CD, CH or CL with lysosomal-associated membrane protein 1 (lamp1) and CD68, the sections were incubated with rabbit anti CB (06-480; Millipore, Temecula, CA, USA) (1:100), CD (Koike et al. 2005; Koike et al. 2013a) (1:100), CH (Kominami et al. 1985) (1:100), or CL (Bando et al. 1986), along with goat anti-lamp1 (AF4800, R&D Systems, Minneapolis, MN) (1:100), and mouse anti-CD68 (1:100).

Techniques: Reflux, Staining, Western Blot

Journal: The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society

Article Title: Increased immunoreactivity of cathepsins in the rat esophagus under chronic acid reflux esophagitis.

doi: 10.1369/0022155414542300

Figure Lengend Snippet: Figure 4. Distribution of cathepsin D (CD) in the esophageal mucosa of sham (A–D) and Reflux Esophagitis (RE) (E–G) groups and the fundus ulcer of the RE group (H–J) at 28 days post-operatively. Triple staining for CD (green), lamp1 (red) and CD68 (blue) in addition to counterstaining with DAPI (white). Boxed areas in C and G are enlarged and shown as insets in C, and H and I, respectively. In the sham group, granular immunoreactivity for CD is intensely localized in the granular layer (g) and absent in the spinous (s) and basal (b) layers (A–C). In the RE group, intense granular staining for CD is observed in the granular layer and weak granular staining appears in the spinous layers, but the immunoreactivity for CD is absent in the basal granular layer (E–G). In the sham group, some CD68-positive macrophages are positive for CD (arrowheads in A–D), whereas some are negative for CD (arrows in A-D). In the RE group, most of the CD68-positive macrophages are intensely positive for CD (H–J). Dotted lines: upper and lower limits of granular and basal layers, respectively. Asterisks: non-specific staining in the lamina propria. c: cornified layer, lp: lamina propria. Scale bars: 20 µm (A–G, J–L), 50 µm (inset in C, H, I). (M) Immunoblot analysis of CD in esophageal mucosae of the sham (S) and RE (R) groups. Mature forms of CD (43 kD and 31 kD for single- and heavy- chain forms, respectively) are increased in the mucosa of the RE group.

Article Snippet: For the triple staining of CB, CD, CH or CL with lysosomal-associated membrane protein 1 (lamp1) and CD68, the sections were incubated with rabbit anti CB (06-480; Millipore, Temecula, CA, USA) (1:100), CD (Koike et al. 2005; Koike et al. 2013a) (1:100), CH (Kominami et al. 1985) (1:100), or CL (Bando et al. 1986), along with goat anti-lamp1 (AF4800, R&D Systems, Minneapolis, MN) (1:100), and mouse anti-CD68 (1:100).

Techniques: Reflux, Staining, Western Blot

Journal: The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society

Article Title: Increased immunoreactivity of cathepsins in the rat esophagus under chronic acid reflux esophagitis.

doi: 10.1369/0022155414542300

Figure Lengend Snippet: Figure 5. Distribution of cathepsin H (CH) in the esophageal mucosa of sham (A–D) and Reflux Esophagitis (RE) (E–G) groups and the fundus ulcer of the RE group (H–J) at 28 days post-operatively. Triple staining for CH (green), lamp1 (red) and CD68 (blue) in addition to counterstaining with DAPI (white). Boxed areas in C and G are enlarged and shown as insets in C, and H and I, respectively. In the sham group, granular immunoreactivity for CH is absent in all layers (A–C). In the RE group, weak but distinct granular staining for CH appears in all layers (E–G), though its immunoreactivity in the basal layer is faint. In both groups, CD68-positive macrophages are intensely positive for CH (arrowheads in A–D and H–J). Dotted lines: upper and lower limits of granular and basal layers, respectively. Asterisks: non- specific staining in the lamina propria. c: cornified layer, g: granular layer, s: spinous layer, b: basal layer, lp: lamina propria. Scale bars: 20 µm (A–G, J–L), 50 µm (inset in C, H, I). (M) Immunoblot analysis of CH in esophageal mucosae of the sham (S) and RE (R) groups. Mature forms of CH (28 kD and 22 kD for single and heavy chains, respectively) are significantly increased in the mucosa of the RE group.

Article Snippet: For the triple staining of CB, CD, CH or CL with lysosomal-associated membrane protein 1 (lamp1) and CD68, the sections were incubated with rabbit anti CB (06-480; Millipore, Temecula, CA, USA) (1:100), CD (Koike et al. 2005; Koike et al. 2013a) (1:100), CH (Kominami et al. 1985) (1:100), or CL (Bando et al. 1986), along with goat anti-lamp1 (AF4800, R&D Systems, Minneapolis, MN) (1:100), and mouse anti-CD68 (1:100).

Techniques: Reflux, Staining, Western Blot

Journal: The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society

Article Title: Increased immunoreactivity of cathepsins in the rat esophagus under chronic acid reflux esophagitis.

doi: 10.1369/0022155414542300

Figure Lengend Snippet: Figure 6. Distribution of cathepsin L (CL) in the esophageal mucosa of sham (A–D) and Reflux Esophagitis (RE) (E–G) groups and the fundus ulcer of the RE group (H–J) at 28 days post-operatively. Triple staining for CL (green), lamp1 (red) and CD68 (blue) in addition to counterstaining with DAPI (white). Boxed areas in C and G are enlarged and shown as insets in C, and H and I, respectively. In the sham group, granular immunoreactivity for CL is weak localized in the granular (g) and basal (b) layers and absent in the spinous (s) layer (A–C). In the RE group, moderate granular staining for CL appears in the basal layer in addition to the weak immunodeposits for CL in the granular layer, but immunoreactivity for CL is also absent in the spinous layers (E–G). In the sham group, some CD68-positive macrophages are positive for CL (arrowheads in A–D), whereas some are negative for CL (arrows in A–D). In the RE group, most of the CD68-positive macrophages are intensely positive for CL (H–J). Dotted lines: upper and lower limits of granular and basal layers, respectively. c: cornified layer, lp: lamina propria. Scale bars: 20 µm (A–G, J–L), 50 µm (an inset in C, H, I). (M) Immunoblot analysis of CL in esophageal mucosae of the sham (S) and RE (R) groups. In both groups, the proform (pro) of CL (39 kD) is intensely detected. Furthermore, the mature forms of CL (29 kD and 23 kD for single and heavy chains, respectively) are increased significantly in the mucosa of the RE group.

Article Snippet: For the triple staining of CB, CD, CH or CL with lysosomal-associated membrane protein 1 (lamp1) and CD68, the sections were incubated with rabbit anti CB (06-480; Millipore, Temecula, CA, USA) (1:100), CD (Koike et al. 2005; Koike et al. 2013a) (1:100), CH (Kominami et al. 1985) (1:100), or CL (Bando et al. 1986), along with goat anti-lamp1 (AF4800, R&D Systems, Minneapolis, MN) (1:100), and mouse anti-CD68 (1:100).

Techniques: Reflux, Staining, Western Blot

Journal: The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society

Article Title: Increased immunoreactivity of cathepsins in the rat esophagus under chronic acid reflux esophagitis.

doi: 10.1369/0022155414542300

Figure Lengend Snippet: Figure 7. Distribution of cathepsin X (CX) in the esophageal mucosa of sham (A–D) and Reflux Esophagitis (RE) (E–G) groups and the fundus ulcer of the RE group (H–J) at 28 days post-operatively. Triple staining for CX (green), lamp1 (red) and CD68 (blue) in addition to counterstaining with DAPI (white). Boxed areas in C and G are enlarged and shown as insets in C, and H and I, respectively. In the sham group, granular immunoreactivity for CX is absent in all layers (A–C). In the RE group, weak but distinct granular staining for CX appears in the basal (b) and spinous (s) layers (E–G), whereas immunoreactivity for CX is absent in the granular layer (g). In both groups, CD68-positive macrophages are positive for CX. Immunoreactivity for CX in macrophages is stronger in the RE groups (arrowheads in A-D and H–J). Dotted lines: upper and lower limits of granular and basal layers, respectively. c: cornified layer, g: granular layer, s: spinous layer, b: basal layer, lp: lamina propria. Scale bars: 20 µm (A–G, J–L), 50 µm (inset in C, H, I). (M) Immunoblot analysis of CX in esophageal mucosae of the sham (S) and RE (R) groups. The proform (pro) of CX (34 kD) is increased significantly in the mucosa of the RE group. The mature form of CX is not detected in both groups.

Article Snippet: For the triple staining of CB, CD, CH or CL with lysosomal-associated membrane protein 1 (lamp1) and CD68, the sections were incubated with rabbit anti CB (06-480; Millipore, Temecula, CA, USA) (1:100), CD (Koike et al. 2005; Koike et al. 2013a) (1:100), CH (Kominami et al. 1985) (1:100), or CL (Bando et al. 1986), along with goat anti-lamp1 (AF4800, R&D Systems, Minneapolis, MN) (1:100), and mouse anti-CD68 (1:100).

Techniques: Reflux, Staining, Western Blot

Journal: The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society

Article Title: Increased immunoreactivity of cathepsins in the rat esophagus under chronic acid reflux esophagitis.

doi: 10.1369/0022155414542300

Figure Lengend Snippet: Figure 8. Distribution of cathepsin F (A– H) and S (I–P) (CF and CS, respectively) in the esophageal mucosa of sham (A–D and I–L, respectively) and Reflux Esophagitis (RE) (E–H and M–P, respectively) groups and the fundus ulcer of the RE group (insets in F, H and N, P, respectively) at 28 days post-operatively. Triple staining for CF or CS (green), lamp1 (red) and CD68 (blue) in addition to counterstaining with DAPI (white). Boxed areas in K and O are enlarged and shown as insets. In both groups, granular immunoreactivity for CF is absent in all layers (A–C and E–G). In both groups, some CD68-positive macrophages are positive for CF (arrowheads in B–D, F–H) and some are negative for CF (arrows in B–D, F–H). Immunoreactivity for CF in macrophages is stronger in the RE groups (arrowheads in A–D, H–J and insets in F, H). Note that granular immunoreactivity for CF is also detected in cells other than macrophages in the lamina propria (lp) and endothelial cells of the capillaries (asterisks). In both groups, granular immunoreactivity for CS is absent in all layers (I–K and M–O). In the sham group, CD68-positive macrophages are completely negative for CS (arrows in J–L), whereas in the RE group CD68-positive macrophages are intensely immunopositive for CS (arrowheads in N–P and insets in O, P). Dotted lines: upper and lower limits of granular and basal layers, respectively. c: cornified layer, g: granular layer, s: spinous layer, b: basal layer. Scale bars: 20 µm (A-P, insets in F, H, N, P), 50 µm (insets in K, O).

Article Snippet: For the triple staining of CB, CD, CH or CL with lysosomal-associated membrane protein 1 (lamp1) and CD68, the sections were incubated with rabbit anti CB (06-480; Millipore, Temecula, CA, USA) (1:100), CD (Koike et al. 2005; Koike et al. 2013a) (1:100), CH (Kominami et al. 1985) (1:100), or CL (Bando et al. 1986), along with goat anti-lamp1 (AF4800, R&D Systems, Minneapolis, MN) (1:100), and mouse anti-CD68 (1:100).

Techniques: Reflux, Staining