Journal: Genes & Development

Article Title: Mice carrying the homologous human shelterin POT1-L259S mutation linked to pulmonary fibrosis show a telomerase deficiency-like phenotype with telomere shortening with increasing mouse generations

doi: 10.1101/gad.352855.125

Figure Lengend Snippet: Degenerative pathologies in Pot1a L261S/L261S knock-in mice. ( A ) The percentage of mice presenting intestinal, lung, and testis degenerative pathologies at 65–75 weeks of age. The number of positive tissues presenting pathological phenotype and the total number of mice analyzed are represented within each bar. Two-sided Fisher's exact test was performed for statistical analysis in each tissue, and the P -values are indicated. ( B ) Representative images of healthy ( top row) and pathological ( middle and bottom rows) intestines, lungs, and testes. The description of the observed pathologies is included below . Two examples of pathologies in each organ are included, and their descriptions are shown below the images. Degenerative intestinal pathologies include epithelial and glandular atrophy with shortened villi and/or decreased number of glands and cystic hyperplasia as well as diffuse inflammatory reaction (black and orange arrows). Degenerative lung pathologies include BALT hyperplasia with hemosiderosis, inflammatory infiltration, and focal thickening of the alveolar septa (yellow and red arrows). Degenerative pathologies in testes include low cellularity and maturation arrest in seminiferous tubules and in some instances focal testicular atrophy with the absence of germinal cells and an increase in Leydig or interstitial cells (green and pink arrows). ( C – E ) Quantification of the percentage of γH2AX in the intestines ( C ) and lungs ( D ), the percentage of Ki67 in the intestines ( E ), and the percentage of smooth muscle actin (SMA) in the lungs ( F ) in 65–75 week old mice of the indicated genotypes. ( G ) Quantification of Masson trichrome-stained lung area in mice of the indicated genotype. Four to six mice were included in each genotype. Statistical significance was addressed by one-way ANOVA with Tukey's multiple comparison test.

Article Snippet: Immunohistochemistry (IHC) was performed on deparaffinized tissue sections processed with 10 mM sodium citrate (pH 6.5) and cooked under pressure for 2 min. IHC staining of tissue sections was performed with phospho-Histone H2AX (ser139) (Millipore 05-636), Ki67 (Cell Signaling Technology 12202), and smooth muscle actin (SMA) (DAKO IR611).

Techniques: Knock-In, Staining, Comparison

Journal: Journal of Histochemistry and Cytochemistry

Article Title: Histomorphological and Molecular Assessments of the Fixation Times Comparing Formalin and Ethanol-Based Fixatives

doi: 10.1369/0022155417741467

Figure Lengend Snippet: Antibodies Used for Immunohistochemistry.

Article Snippet: Positive controls included human colon, spleen, and kidney for Histone H3, Ki-67 and AQP1, respectively. table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Antibody Vendor Clonality Cat. No. Incubation Dilution Antigen Retrieval AQP1 Santa Cruz Biotechnology (Dallas, TX) Poly rabbit Sc-20810 30 min at RT 1:250 Not applied Histone H3 Abcam (Cambridge, MA) Poly rabbit ab70550 30 min at RT 1:2000 20 min, pressure cooker, pH 6 Ki-67 Sigma-Aldrich (St. Louis, MO) Mono rabbit (SP6) 275R 30 min at RT Ready to use 20 min, pressure cooker, pH 6 Open in a separate window Abbreviations: AQP1, aquaporin 1; RT, room temperature.

Techniques: Immunohistochemistry, Incubation

Journal: Journal of Histochemistry and Cytochemistry

Article Title: Histomorphological and Molecular Assessments of the Fixation Times Comparing Formalin and Ethanol-Based Fixatives

doi: 10.1369/0022155417741467

Figure Lengend Snippet: Immunohistochemical assessment for the fixatives BE70 and NBF according to the fixation time. Tissue sections were reacted with rabbit polyclonal Histone H3 antibodies. (A) Representative images of IHC staining of Histone H3 expression in mouse spleen tissue. Scale bar, 50 µm. (B) IHC images were analyzed using digital image analyses. The right panel shows the relative Histone H3 expressional signal according to the fixation time in the BE70 or NBF fixatives. The bar graph shows the relative averages of the histoscores. The relative Histone H3 expressional signal of each entity was normalized to 1 day BE70-fixed tissue (1.00). Abbreviations: BE70, buffered ethanol; NBF, neutral buffered formalin; AQP1, aquaporin 1. *p<0.05, **p<0.01, ***p<0.001.

Article Snippet: Positive controls included human colon, spleen, and kidney for Histone H3, Ki-67 and AQP1, respectively. table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Antibody Vendor Clonality Cat. No. Incubation Dilution Antigen Retrieval AQP1 Santa Cruz Biotechnology (Dallas, TX) Poly rabbit Sc-20810 30 min at RT 1:250 Not applied Histone H3 Abcam (Cambridge, MA) Poly rabbit ab70550 30 min at RT 1:2000 20 min, pressure cooker, pH 6 Ki-67 Sigma-Aldrich (St. Louis, MO) Mono rabbit (SP6) 275R 30 min at RT Ready to use 20 min, pressure cooker, pH 6 Open in a separate window Abbreviations: AQP1, aquaporin 1; RT, room temperature.

Techniques: Immunohistochemical staining, Immunohistochemistry, Expressing