Figure 5.
Figure 5. Analyses of femur BM sections, spleen weight, and SLAMF7 expression of monocytes in the spleen and PB of hNOG mice treated with Rom and Elo. (A) The protocol for the transplantation of human cord blood into NOG mice and the administration of Rom and Elo to transplanted hNOG mice in this study. (B) The silver staining and immunohistochemical staining with α-smooth muscle actin (α-SMA) of a femur BM section from hNOG mice treated with Rom and Elo. Both the silver-stained and α-SMA-stained BM sections showed increased fibrosis. (Left) The BM section of mice treated with Rom alone. The BM stroma with silver staining exhibited dense reticulin fibers intermingled with bundles of collagen. In addition, the BM stroma was diffusely α-SMA positive and stained brown in α-SMA staining. MF grade was evaluated as MF-3. The image is representative of 10 mice treated with Rom alone. (Right) The BM section of mice treated with Rom and Elo. The BM stroma exhibited focally dense reticulin fibers forming extensive intersections and was partially α-SMA positive. MF grade was evaluated as MF-2. Elo suppressed Rom-induced MF. The image is representative of 12 mice treated with Rom plus Elo. Pictures were taken and digitized by a BZ-X700 microscope and with BZ-H3A software. Original magnification ×20 for panel B. Scale bar, 300 μm. (C) The spleen weight of the treated hNOG mice. Both mice treated with Rom alone (n = 10) and mice treated with Rom plus Elo (n = 12) exhibited higher spleen weight than control mice (n = 5). The spleen weight significantly decreased in mice treated with Rom plus Elo compared with mice treated with Rom alone. (D) Analysis of the relative rate of SLAMF7 expression (anti-SLAMF7 antibody/isotype control) of CD45+CD14+ monocytes in the spleen of the treated hNOG mice. The relative rate of SLAMF7 expression of monocytes in the spleen significantly increased in mice treated with Rom alone (n = 10) compared with control mice (n = 5) and mice treated with Rom plus Elo (n = 12). (E) Analysis of the relative rate of SLAMF7 expression (anti-SLAMF7 antibody/isotype control) of CD45+CD14+ monocytes in the PB of the treated hNOG mice. The relative rate of SLAMF7 expression in monocytes in PB significantly increased in mice treated with Rom alone (n = 10) compared with control mice (n = 5) and mice treated with Rom plus Elo (n = 12). (F) Analysis of the relative rate of SLAMF7 expression (anti-SLAMF7 antibody/isotype control) in lymphocytes in the spleen of treated hNOG mice. No significant changes were observed in the relative rate of SLAMF7 expression of lymphocytes in the spleen among control mice (n = 5), mice treated with Rom alone (n = 10), and mice treated with Rom plus Elo (n = 12). (G) Analysis of the relative rate of SLAMF7 expression (anti-SLAMF7 antibody/isotype control) in lymphocytes in PB of treated hNOG mice. No significant changes were observed in the relative rate of SLAMF7 expression of lymphocytes in PB among control mice (n = 5), mice treated with Rom alone (n = 10), and mice treated with Rom plus Elo (n = 12), which is the same as the lymphocytes in the spleen. Results are reported as mean ± standard deviation values. Statistical significance was calculated using 1-way ANOVA with post hoc Tukey’s honestly significant difference test. *P < .05; **P < .01.

Analyses of femur BM sections, spleen weight, and SLAMF7 expression of monocytes in the spleen and PB of hNOG mice treated with Rom and Elo. (A) The protocol for the transplantation of human cord blood into NOG mice and the administration of Rom and Elo to transplanted hNOG mice in this study. (B) The silver staining and immunohistochemical staining with α-smooth muscle actin (α-SMA) of a femur BM section from hNOG mice treated with Rom and Elo. Both the silver-stained and α-SMA-stained BM sections showed increased fibrosis. (Left) The BM section of mice treated with Rom alone. The BM stroma with silver staining exhibited dense reticulin fibers intermingled with bundles of collagen. In addition, the BM stroma was diffusely α-SMA positive and stained brown in α-SMA staining. MF grade was evaluated as MF-3. The image is representative of 10 mice treated with Rom alone. (Right) The BM section of mice treated with Rom and Elo. The BM stroma exhibited focally dense reticulin fibers forming extensive intersections and was partially α-SMA positive. MF grade was evaluated as MF-2. Elo suppressed Rom-induced MF. The image is representative of 12 mice treated with Rom plus Elo. Pictures were taken and digitized by a BZ-X700 microscope and with BZ-H3A software. Original magnification ×20 for panel B. Scale bar, 300 μm. (C) The spleen weight of the treated hNOG mice. Both mice treated with Rom alone (n = 10) and mice treated with Rom plus Elo (n = 12) exhibited higher spleen weight than control mice (n = 5). The spleen weight significantly decreased in mice treated with Rom plus Elo compared with mice treated with Rom alone. (D) Analysis of the relative rate of SLAMF7 expression (anti-SLAMF7 antibody/isotype control) of CD45+CD14+ monocytes in the spleen of the treated hNOG mice. The relative rate of SLAMF7 expression of monocytes in the spleen significantly increased in mice treated with Rom alone (n = 10) compared with control mice (n = 5) and mice treated with Rom plus Elo (n = 12). (E) Analysis of the relative rate of SLAMF7 expression (anti-SLAMF7 antibody/isotype control) of CD45+CD14+ monocytes in the PB of the treated hNOG mice. The relative rate of SLAMF7 expression in monocytes in PB significantly increased in mice treated with Rom alone (n = 10) compared with control mice (n = 5) and mice treated with Rom plus Elo (n = 12). (F) Analysis of the relative rate of SLAMF7 expression (anti-SLAMF7 antibody/isotype control) in lymphocytes in the spleen of treated hNOG mice. No significant changes were observed in the relative rate of SLAMF7 expression of lymphocytes in the spleen among control mice (n = 5), mice treated with Rom alone (n = 10), and mice treated with Rom plus Elo (n = 12). (G) Analysis of the relative rate of SLAMF7 expression (anti-SLAMF7 antibody/isotype control) in lymphocytes in PB of treated hNOG mice. No significant changes were observed in the relative rate of SLAMF7 expression of lymphocytes in PB among control mice (n = 5), mice treated with Rom alone (n = 10), and mice treated with Rom plus Elo (n = 12), which is the same as the lymphocytes in the spleen. Results are reported as mean ± standard deviation values. Statistical significance was calculated using 1-way ANOVA with post hoc Tukey’s honestly significant difference test. *P < .05; **P < .01.

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