Journal: Cancer science

Article Title: Coordinate suppression of Sdpr and Fhl1 expression in tumors of the breast, kidney, and prostate.

doi: 10.1111/j.1349-7006.2008.00816.x

Figure Lengend Snippet: Fig. 1. Fhl1 and Sdpr expression in normal epi- thelium and tumors from human breast, kidney, and prostate. Immunohistochemical staining was performed on normal and tumor human tissue. Fhl1 and Sdpr staining was evident in normal epithelial cells of the breast, kidney, and prostate. In contrast, both Fhl1 and Sdpr expression were suppressed in infiltrating mammary duct carcinoma, renal cell carcinoma, and prostate adenocarcinoma. (bar = 60 microns).

Article Snippet: Slides containing 15 infiltrating duct carcinoma specimens with normal adjacent tissue from breast cancer patients (IMH-371, Imgenex), nine prostate adenocarcinoma specimens with adjacent normal tissue and an additional unmatched 41 prostate adenocarcinoma specimens (IMH-303, Imgenex), and nine renal cell carcinoma specimens with adjacent normal tissue and an additional unmatched 31 cell carcinoma specimens (IMH-313, Imgenex) were examined by immunohistochemistry as previously described. (19) Briefly, after removal of paraffin with xylene and rehydration through a graded series of decreasing concentrations of ethanol, protein antigenicity was enhanced by microwaving sections in Target Buffer (Dako, Carpenturia, CA, USA) for 2 min.

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: Cell Death Discovery

Article Title: Enhanced apoptosis as a possible mechanism to self-limit SARS-CoV-2 replication in porcine primary respiratory epithelial cells in contrast to human cells

doi: 10.1038/s41420-021-00781-w

Figure Lengend Snippet: A – D Formalin-fixed paraffin-embedded cross-sections of human and pig tracheal sections stained for ACE2 with ImmPRESS VR anti-mouse IgG horseradish peroxidase (HRP) polymer detection kit (MP-6402-15). Brown spots represent the presence of ACE2 receptor that interacted with specific antibody is considered a positive expression, and the nucleus counterstained with hematoxylin is blue. Expression of ACE2 in human ( A ) and pig ( C ) trachea revealed by immunostaining with mouse anti-ACE2 monoclonal antibody (4 μg/mL; sc-390851). Note the focal expression of ACE2 on the epithelial cells of pig trachea ( C ; arrow in inset image). B , D Corresponding negative control tissues sections stained with secondary anti-mouse HRP antibody only. Scale bar-100 μm (inset 25 μm). Pig ( n = 3); human ( n = 1). Microscopic morphology of primary human ( E ; HRECs) and pig ( F ; PRECs) respiratory epithelial cells. G Determination of pan-cytokeratin and ACE2 in HRECs and PRECs using flow cytometry. Cells stained with LIVE/DEAD ® Fixable Near-IR Dead Cell Stain Kit, and respective mouse monoclonal antibodies (epithelial cell marker pan-cytokeratin; 0.5 μg/mL; MCA1907T); (ACE2; 4 μg/mL; sc-390851,) incubated for 30 min followed by incubation with Alexa Fluor ® 647 Goat anti-mouse (15 μg/mL) for another 30 min. Data was collected using an Attune NxT flow cytometer. A representative of 10,000 events were acquired and analyzed for each sample. Cells were gated for singlet population using forward (FSA) and side-scatter (SSA) properties, and the mean of percent live cell population was used to quantify the levels of pan-cytokeratin and ACE2 ( n = 4). The bar graph represents the mean (SD).

Article Snippet: Normal human trachea tissue slides were commercially bought (Novus, NBP2-77809 Novus Biologicals, LLC, Centennial, CO, USA), while pig trachea sections were collected for this study.

Techniques: Formalin-fixed Paraffin-Embedded, Staining, Polymer, Expressing, Immunostaining, Negative Control, Flow Cytometry, Marker, Incubation

Journal: Cell Death Discovery

Article Title: Enhanced apoptosis as a possible mechanism to self-limit SARS-CoV-2 replication in porcine primary respiratory epithelial cells in contrast to human cells

doi: 10.1038/s41420-021-00781-w

Figure Lengend Snippet: The tissues and cells presented in this panel were stained with ImmPRESS VR anti-rabbit IgG horseradish peroxidase (HRP) polymer detection kit (MP-6401-15) with a rabbit monoclonal antibody specific for anti-SARS Coronavirus/SARS-Related Coronavirus 2 nucleocapsid (N) protein (0.75 μg/mL) (the following reagent was obtained through BEI Resources, NR-53791). Dark brown spots represent a positive expression of viral N protein, and pale brown represents background staining, while the nucleus counterstained with hematoxylin is blue. A – D Representative images showing SARS-CoV-2 binding in formalin-fixed paraffin-embedded cross-sections of A human and B pig trachea after overnight incubation with 250 μL heat-inactivated SARS-Related Coronavirus 2, Isolate USA-WA1/2020 (the following reagent was obtained through BEI Resources; NR-52286). C Human ( n = 1) and D pig ( n = 3) tracheal epithelial sections were stained with secondary anti-rabbit HRP antibody only. E – P Detection of SARS-CoV-2 infection in HRECs and PRECs inoculated with SARS-CoV-2, Isolate USA-WA1/2020 at 120 hpi. Paraformaldehyde (4%) fixed cells were stained for SARS-CoV-2 N protein. E , I , and M HRECs and F , J , and N PRECS were inoculated with a viral dose of MOI 5.0 or culture medium, as negative control ( G , K , and O for HRECs; H , L , and P for PRECs); ( n = 6). Scale bar—100 μm.

Article Snippet: Normal human trachea tissue slides were commercially bought (Novus, NBP2-77809 Novus Biologicals, LLC, Centennial, CO, USA), while pig trachea sections were collected for this study.

Techniques: Staining, Polymer, Expressing, Binding Assay, Formalin-fixed Paraffin-Embedded, Incubation, Infection, Negative Control

Journal: Hypertension

Article Title: Reduced Notch1 Cleavage Promotes the Development of Pulmonary Hypertension

doi: 10.1161/hypertensionaha.120.16065

Figure Lengend Snippet: Figure 3. Enhanced endothelial cell (EC) permeability and inflammation by Delta-like 4 neutralizing antibody (Dll4nAb) treatment in normal mice lung tissues. A, En face staining of N1-ICD (red) and VE-cadherin (green) on ECs of pulmonary arteries of Dll4nAb injected mice. Arrows, impaired adherent junction integrity (n=3 per group). Nuclei were counterstained by DAPI (blue). Scale bars, 10 µm. B, Quantification of trans- endothelial electrical resistance (TEER) and (C) paracellular permeability in confluent cultures of adult human pulmonary microvascular endothelial cell monolayers in the absence or presence of DAPT (10 µM) treatment for 12 h (n=4). (Continued )

Article Snippet: Normal lung tissue slides were purchased from Novus Biologicals (NBP2-30182).

Techniques: Permeability, Staining, Injection