Figure 3.
Spleen histology, BM neutrophil content and hematopoietic progenitors in female Dnm2het mice. (A) Comparison of spleen weights in female Dnm2het and CTRL mice. Overall, when comparing groups involving all ages (N = 18 and above), female Dnm2het mice developed splenomegaly. (B) Spleen histology in 12- to 25-week-old female Dnm2het and CTRL mice, showing florid reactive germinal center hyperplasia with the expansion of BCL-6 positive lymphocytes within secondary reactive follicles. CTRL mice: sections demonstrate a normal proportion and white and red pulp distribution. BCL-6 immunostain shows the absence of secondary reactive follicles. Dnm2het mice: the white pulp was mildly expanded by reactive, secondary follicles. BCL-6 highlights germinal center lymphocytes. Objective: 10×. (C) Average percentage of Ly6G positive BM cells in female Dnm2het and CTRL mice when comparing groups involving all ages (N = 24 and above). Scoring by immunohistochemistry demonstrated decreased BM Ly6G positive neutrophil. Histology slide images were captured using the Aperio AT2 slide scanner at 20X or the CRI Nuance Spectral Imaging microscope, unless specified otherwise. Quantification was performed by Halo Image analysis. (D) Average total GMP (lineage negative, R-IL7 negative, c-Kit positive, Sca-1 negative, CD34 positive, CD16/32 high) in the BM of female Dnm2het and CTRL mice when comparing groups involving all ages (N = 17 and above). (E) Average total BM lineage negative (Lin neg) cells in the BM of female Dnm2het and CTRL mice (N = 23 and above). (F) Average total BM lineage negative, R-IL7 negative, Sca-1 positive, c-Kit positive (LSK) cells in the BM of female Dnm2het and CTRL mice when comparing groups involving all ages (N = 18 and above). Dnm2het female mice displayed a decrease in BM Lin neg cells counted after immunomagnetic lineage depletion, decreased LSK and GMP measured by flow cytometry when comparing groups involving all ages. (G) Representative plot of LSK BM cells flow cytometry analysis. (H) Representative plot of megakaryocyte-erythrocyte progenitor (MEP), common myeloid progenitor (CMP), and GMP flow cytometry analysis. Data are shown as mean +/− SEM. ∗P < .05, ∗∗P < .01, t-test.

Spleen histology, BM neutrophil content and hematopoietic progenitors in female Dnm2het mice. (A) Comparison of spleen weights in female Dnm2het and CTRL mice. Overall, when comparing groups involving all ages (N = 18 and above), female Dnm2het mice developed splenomegaly. (B) Spleen histology in 12- to 25-week-old female Dnm2het and CTRL mice, showing florid reactive germinal center hyperplasia with the expansion of BCL-6 positive lymphocytes within secondary reactive follicles. CTRL mice: sections demonstrate a normal proportion and white and red pulp distribution. BCL-6 immunostain shows the absence of secondary reactive follicles. Dnm2het mice: the white pulp was mildly expanded by reactive, secondary follicles. BCL-6 highlights germinal center lymphocytes. Objective: 10×. (C) Average percentage of Ly6G positive BM cells in female Dnm2het and CTRL mice when comparing groups involving all ages (N = 24 and above). Scoring by immunohistochemistry demonstrated decreased BM Ly6G positive neutrophil. Histology slide images were captured using the Aperio AT2 slide scanner at 20X or the CRI Nuance Spectral Imaging microscope, unless specified otherwise. Quantification was performed by Halo Image analysis. (D) Average total GMP (lineage negative, R-IL7 negative, c-Kit positive, Sca-1 negative, CD34 positive, CD16/32 high) in the BM of female Dnm2het and CTRL mice when comparing groups involving all ages (N = 17 and above). (E) Average total BM lineage negative (Lin neg) cells in the BM of female Dnm2het and CTRL mice (N = 23 and above). (F) Average total BM lineage negative, R-IL7 negative, Sca-1 positive, c-Kit positive (LSK) cells in the BM of female Dnm2het and CTRL mice when comparing groups involving all ages (N = 18 and above). Dnm2het female mice displayed a decrease in BM Lin neg cells counted after immunomagnetic lineage depletion, decreased LSK and GMP measured by flow cytometry when comparing groups involving all ages. (G) Representative plot of LSK BM cells flow cytometry analysis. (H) Representative plot of megakaryocyte-erythrocyte progenitor (MEP), common myeloid progenitor (CMP), and GMP flow cytometry analysis. Data are shown as mean +/− SEM. ∗P < .05, ∗∗P < .01, t-test.

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