BM dysplastic feature analysis and autophagy marker LC3B staining pattern in older female Dnm2^het mice. (A) BM dysplastic morphology analysis in female and Dnm2^het CTRL mice. After 50 weeks of age, 26% of Dnm2^het mice (out of N = 23), but no CTRL mice, displayed marrow dysplasia. On hematoxylin & eosin stain, with red arrows indicating megakaryocytes, Dnm2^het mice show frequent dysmorphic megakaryocytes with small and hyposegmented nuclei with a scant amount of cytoplasm, whereas CTRL mice showed normal segmented megakaryocytes with variable amounts of cytoplasm. Histology slide images were captured using the Aperio AT2 slide scanner at 20X or the CRI Nuance Spectral Imaging microscope, unless specified otherwise. (B) Autophagy marker LC3B pattern in older female Dnm2^het mice (>34 weeks of age). For imaging of LC3B cluster dots, we used an Olympus BH2 microscope with a 100X objective, NA 1.3, and Olympus C-35AD-2 camera. Additional nonlinear adjustments such as changes to gamma settings using the Photoshop software were applied to improve discernibility on the digital images. Autophagy marker LC3B stained dotted clusters were more conspicuous and more readily observed in the cytoplasm in the BM of young CTRL (<34 weeks of age) than in the BM in the other groups (old CTRL above 34 weeks combined with Dnm2^het all ages). (C,D) Halo Image analysis of the average cytoplasm LC3B staining in different mouse groups. The average cytoplasm LC3B staining was lower in the young CTRL (<34 weeks) than in other groups taken together (old CTRL above 34 weeks plus Dnm2^het all ages), N = 8 and above. (E) Halo Image analysis of the average nuclear size in young Dnm2^het females (<34 weeks) than in young CTRL females (<34 weeks). The average nuclear size was greater in young Dnm2^het females than in young CTRL females, N = 4 and above. Data are shown as mean +/− SEM. ∗P < .05, ∗∗P < .01, t-test.