Journal: Animal Models and Experimental Medicine

Article Title: Initial joint bleed volume in a delayed on‐demand treatment setup correlates with subsequent synovial changes in hemophilic mice

doi: 10.1002/ame2.12118

Figure Lengend Snippet: Histopathological evaluation of knees 14 days after induced joint bleed. A, Representative histological images of injured knees (aʹ, bʹ) and 1 contralateral knee displaying spontaneous hemophilic arthropathy (cʹ). First column: Hematoxylin eosin (HE) stain at 20 × magnification. Blue rectangle shows the area magnified in the second column, whereas the white rectangle shows the area magnified in columns 3‐5, all at 400 × magnification. Second column: HE of tibial cartilage, showing no loss of chondrocytes or cartilage erosion. Third column: HE of the synovium, with hyperplasia present in bʹ and cʹ. Fourth column: Perl's Prussian blue stain; arrows indicate hemosiderin deposition. Fifth column: α‐sma stain, arrows mark blood vessels. B, Synovitis score in injured and contralateral knees (mean ± SD). C, Vessel count in the synovium along the femur in injured and contralateral knees. D, Area of hemosiderin (Perls area) in injured and contralateral knees. E, Chondrocyte in injured and contralateral knees. Significance levels are marked as follows: * P < .05, **** P < .0001

Article Snippet: Perls’ Prussian Blue stain was used to quantify the hemosiderin‐stained area in the joint (outside the bone marrow) using automatic image analysis (VIS 2019.02.1.6005; Visiopharm).

Techniques: H&E Stain, Staining

Journal: Life Science Alliance

Article Title: Redundant and nonredundant organismal functions of EPS15 and EPS15L1

doi: 10.26508/lsa.201800273

Figure Lengend Snippet: (A) Methylene blue–stained blood smears of adult WT mice, 7 d after injection with PBS or PHZ. Note numerous blue dye–retaining reticulocytes in the blood smear from PHZ-treated mice. Bar, 10 μm. (B) FACS analysis with anti-TfR/CD71 and thiazole orange to analyze the maturation status of RBCs from WT mice at 0, 7, 10, and 16 d after PHZ injection. (C) Western blotting of blood lysates at 7, 10, and 16 d after treatment with PHZ. Equal amounts of lysates, corresponding to 7 μl of whole blood, were loaded and probed for the indicated proteins. (D) Western blotting of brain (Br), liver (Li), and spleen (Sp) lysates from adult WT and cDKO mice. Twenty micrograms of each lysate was loaded. (E) Iron, transferrin, and ferritin levels in the serum of adult WT and cDKO mice. Three animals of each phenotype were analyzed. ** P < 0.01 versus WT. Note that the iron levels were increased in the serum of cDKO mice, indicating that iron absorption was not defective. (F) Perls’ Prussian blue staining of the liver and spleen of adult WT and cDKO mice. Iron deposition is in blue. Arrows point to increased erythropoiesis in the spleen of cDKO mice. Bar, 50 μm. Note that, despite increased serum iron, we did not detect tissue iron overload in the spleen or liver of cDKO mice. In contrast, other murine models of microcytic hypochromic anemia, such as the hematopoietic-specific KO for Stat5a/b , present iron overload in the liver . A possible explanation might be that cDKO displayed increased secondary erythropoiesis, particularly in the spleen (this panel), which actively remove iron excess. Consistent with the absence of tissue iron overload, other iron metabolism proteins (transferrin and ferritin) were not altered in cDKO mice (panel E), indicating that a certain balance in iron metabolism in cDKO mice has still been preserved and/or re-established. (G) Molecular phylogenesis of the EPS15 family. Protein sequences were retrieved from the NCBI or the Joint Genome Institute ( http://genome.jgi.doe.gov/ ) databases. Sequences were aligned with ClustalW, and the evolutionary history was inferred by using the maximum likelihood method based on the JTT matrix–based model . The tree is drawn to scale, with branch lengths measured in the number of substitutions per site. All positions containing gaps and missing data were eliminated. Alignment and evolutionary analyses were conducted in MEGA7 . Protein accession numbers are shown.

Article Snippet: Staining was performed using Perls' Prussian blue stain kit (DDK Italia), according to the manufacturer's instructions.

Techniques: Staining, Injection, Western Blot