Journal: Cell death discovery

Article Title: Inhibition of Aurora Kinase B activity disrupts development and differentiation of salivary glands.

doi: 10.1038/s41420-020-00393-w

Figure Lengend Snippet: Fig. 8 AURKB loss disrupted acinar identity via ROS generation. A, B Mist1 (green) and pan cytokeratin (blue) expressions in E16.5+24h bud cells, scale bar = 50 µm. Remarkable loss of the acinar differentiation marker Mist1 at the protein and transcriptional levels in response to Barasertib treatment. Recovery of Mist1 levels due to ROS scavenging using aminoguanidine (number of pooled glands ≥5), analysed by ANOVA **P ≤0.01, *p ≤0.05, ****P ≤0.0001. C, D ROS production in response to AURKB inhibition in the embryonic salivary glands. ROS levels were visualised in the SMG explants after staining with DCFDA and inspection under fluorescence microscope. Analysis of corrected total fluorescence (CTF) using ImageJ revealed that Barasertib treatment for 24 h significantly increased levels of ROS in the E16.5+24h explants which were significantly reduced after co- treating the explants with aminoguanidine, analysed by ANOVA, **P ≤0.01, *p ≤0.05, ns: non-significant, p > 0.05. Scale bar = 500 μm, n = 3/group.

Article Snippet: To block endogenous peroxidase activity and nonspecific background staining sections were incubated in 3% hydrogen peroxide solution for 20–30 min. To block all epitopes on the tissue samples and prevent nonspecific antibody binding, sections were incubated with 1% BSA in 1X TBS, pH 7.6 for 5 min. Tissue sections were incubated at 4 °C overnight, with the antibody: rabbit-anti-cleaved caspase 3 (1:2500, NB100–56113, Novus Bio), rabbit–anti-Mist1 (1:200, ab187978, Abcam), mouse-anti-Cyclin D1 (1:400, NBP2–32840, Novus Bio), rabbit-anti-AURKB (1:200, ab115793, Abcam), rabbit-anti-ki67 (1:100, ab16667, Abcam), rabbit-anti-p21 (1:200, ab188224, Abcam), rabbit-anti-p.H2AX(1:200, ab81299, Abcam), rat-anti-E- Cadherin (1:200, sc-59778, Santa Cruz Biotechnology, Inc.) guinea pig-anti-pan cytokeratin (1:50, BP5069, Origene).

Techniques: Marker, Inhibition, Staining, Microscopy

Journal: Cancer Management and Research

Article Title: Primary clear cell carcinoma of the thymus and literature comparison of features

doi: 10.2147/cmar.s158452

Figure Lengend Snippet: Figure 2 (A) Microscopic appearance of the surgical specimen. Large lobules were composed of tumor cells closely spaced with fibrotic stroma of varying abundance and density (×40). Bar, 250 μm. (B) Uniform neoplastic cells with abundant, clear cytoplasm, nuclear atypia was not marked, and nuclear chromatin was finely dispersed, nucleoli were not prominent (×400). Bar, 25 μm. (C) Hyaline material is PAS-positive (×200). Bar, 50 μm. The neoplastic cells were positive for pan-cytokeratin (CKpan). (D) (×200), p63, (E) (×200), 34βE12, (F) (×200). Bar, 50 μm. In contrast, neoplastic cells were negative for TTF-1 (G) (×200), GATA-3 (H) (×200), HMB45 (I) (×200). Bar, 50 μm. Abbreviations: PAS, Periodic acid Schiff; TTF-1, thyroid transcription factor 1; GATA-3, GATA binding protein 3; HMB45, human melanoma black 45.

Article Snippet: Immunohistochemical staining was performed using antibodies against pan-cytokeratin (CKpan) (1:100; Beijing Zhongshan Golden Bridge Biotechnology Co., Ltd, Beijing, China), 34βE12 (1:100; Zsbio), p63 (1:100, Zsbio), thyroid transcription factor 1 (TTF-1) (1:25, Zsbio), GATA binding protein 3 (GATA-3) (1:100, Zsbio), human melanoma black 45 (HMB45) (1:100, Zsbio), CD5 (1:100; Zsbio), c-kit (1:50; Zsbio), and terminal deoxynucleotidyl transferase (TdT) (1:50; Zsbio).

Techniques: Binding Assay

Journal: American Journal of Physiology - Renal Physiology

Article Title: Urinary extracellular vesicle-associated MCP-1 and NGAL derived from specific nephron segments differ between calcium oxalate stone formers and controls

doi: 10.1152/ajprenal.00515.2018

Figure Lengend Snippet: Numbers of MCP-1-, NGAL-, and OPN-associated urinary extracellular vesicles from the different segmental epithelium of nephrons in nonstone formers (controls) and idiopathic calcium stone formers

Article Snippet: Phycoerythrin (PE)-conjugated mouse anti-human MCP-1 (catalog no. 502604, BioLegend, San Diego, CA), rabbit anti-human NGAL (catalog no. bs-0444R-PE, Bioss Antibodies, Woburn, MA), mouse anti-human OPN (catalog no. IC14331P, R&D Systems, Minneapolis, MN), FITC-conjugated recombinant annexin V (marker for vesicles carrying surface phosphatidylserine, catalog no. 556419, BD Pharminagen, San Jose, CA), rabbit anti-human aquaporin-1 (marker of the simple squamous epithelium of the descending limb of Henle, catalog no. orb15121, Biorbyt, Cambridge, UK), rabbit anti-human aquaporin-2 (marker of principal cells of the collecting duct, catalog no. orb13806, Biorbyt), rabbit polyclonal anti-human urate transporter (URAT)1 (marker of the simple cuboidal epithelium of the proximal convoluted tubule, catalog no. bs-10357R-FITC, Bioss Antibodies], mouse anti-human uromodulin (marker of the thick ascending loop of Henle, catalog no. LS-B3105, Lifespan Biosciences, Seattle, WA), rabbit polyclonal anti-human SLC12A3 (marker of the simple cuboidal epithelium of the distal convoluted tubule, catalog no. bs-7694R-FITC, Bioss Antibodies), and rabbit anti-human cytokeratin-19 (marker of the transitional epithelium of the papillary duct, catalog no. bs-2190R-FITC, Bioss Antibodies) antibodies were used in this study.

Techniques: Marker, Biomarker Assay