rabbit anti ace2  (Novus Biologicals)

Journal: Scientific Reports

Article Title: Impact of in vitro SARS-CoV-2 infection on breast cancer cells

doi: 10.1038/s41598-024-63804-3

Figure Lengend Snippet: SARS-CoV-2 replication in breast cancer cell lines. Western blot and densitometric analysis of ( a ) ACE2 and ( b ) NRP1 protein expression in the MCF7, MDA-MB-231 and HCC1937 breast cancer cell lines. ( c ) ACE2 and NRP1 were silenced by siRNAs in the MCF7, MDA-MB-231 and HCC1937 breast cancer cell lines, which were subsequently infected with SARS-CoV-2 lineage B1. Viral RNA, expressed as copies/100 ng RNA, was quantified in the cytoplasm 6 h p.i. by real-time PCR with primers targeting the viral N1 gene. The data are presented as the means ± SEMs and are representative of one of three independent experiments with similar results. * p < 0.05 by one-way ANOVA followed by Tukey’s multiple comparison test with a single pooled variance. ( d ) MCF7, MDA-MB-231 and HCC1937 breast cancer cells were infected with SARS-CoV-2, and viral RNA was quantified at different time points in the cytoplasm and in the supernatants by real-time PCR. The viral load is expressed as copies/100 ng RNA or copies/mL for the cellular or supernatant RNA, respectively. The data are presented as the means ± SEMs and are representative of one of three independent experiments with similar results. *** p < 0.001 according to two-way ANOVA with multiple comparisons; refers to the comparison between MCF7 cells and MDA-MB-231 cells and the comparison between MCF7 cells and HCC1937 cells.

Article Snippet: The recombinant rabbit monoclonal anti-ACE2 antibody (clone SN0754, dilution 1:1000, Thermo Fisher Scientific, Waltham, MA, USA), rabbit monoclonal anti-Neuropilin-1 (NRP1) antibody (clone D62C6, dilution 1:1000, Cell Signaling Technology Inc., Danvers, MA, USA) and horseradish peroxidase-conjugated mouse monoclonal anti-β-Actin antibody (clone AC-15, dilution 1:30,000, Merck KGaA, Darmstadt, Germany) served as primary antibodies.

Techniques: Western Blot, Expressing, Infection, Real-time Polymerase Chain Reaction, Comparison

Journal: Viruses

Article Title: Histopathological and Immunological Findings in the Common Marmoset Following Exposure to Aerosolized SARS-CoV-2

doi: 10.3390/v14071580

Figure Lengend Snippet: Representative images of IHC for ACE2 receptor in marmoset tissues. Positive staining was observed in the small intestine (Bar = 250 µm) ( A ), mostly in the apical border of absorptive epithelial cells and at a low level in some epithelial cells of glands. Minimal staining in epithelial cells from the large intestine (Bar = 100 µm) ( B ). The positive staining in the trachea (Bar = 100 µm) ( C ), lung (Bar = 250 µm) ( D ), and respiratory nasal mucosa (Bar = 100 µm) ( E ) was almost non-existent. However, moderate positive staining was observed in glandular epithelium within the olfactory mucosa area of the nasal cavity (arrows and insert; Bar = 250 µm) ( F ). To the contrary, there was no staining in the epithelium lining or nerve terminations. Intense staining was observed in the kidney in the apical border of kidney tubular epithelial cells (arrows; Bar = 250 µm) ( G ) and moderate reaction in immune cells within the liver (Bar = 100 µm; arrows, normally extramedullary haematopoiesis) ( H ). No reaction was observed in the spleen (Bar = 250 µm) ( I ).

Article Snippet: Anti-ACE2 rabbit monoclonal antibody (NBP267692, Novus Biologicals, Centennial, CO, USA) and TMPRSS2 rabbit polyclonal antibody (NBP293322, Novus Biologicals) were used at dilutions of 1:250 and 1:1000, respectively, and were incubated for 30 min. Leica polymer refine detection kit (DS9800, Leica Biosystems) was used, and DAB as chromogen for visualization and sections were counterstained with Harris’ haematoxylin (DS9800, Leica Biosystems).

Techniques: Staining

Journal: bioRxiv

Article Title: Sequential infection with influenza A virus followed by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) leads to more severe disease and encephalitis in a mouse model of COVID-19

doi: 10.1101/2020.10.13.334532

Figure Lengend Snippet: ACE2 expression in the lungs. A. Wild type mouse, uninfected. The epithelium lining the bronchioles (B) shows ACE2 expression at the luminal surface (inset). B. Mock infected K18-hACE2 mouse. Besides the epithelium lining the bronchioles (B), there are scattered type II pneumocytes that express ACE2 (insets: arrowheads). C. K18-hACE2 mouse, 3 days post SARS-CoV-2 infection. The expression pattern and extent is comparable to that in the mock infected K18-hACE2 mouse. Arrowheads point at ACE2 positive type II pneumocytes. D. K18-hACE2 mouse, 7 days post SARS-CoV-2 infection. The expression pattern is identical to that in the mock infected K18-hACE2 mouse, but activated type II pneumocytes that express ACE2 (arrowheads) are more numerous. E. K18-hACE2 mouse, 10 days post IAV infection. In unaltered bronchioles, the ACE2 expression pattern is identical to that in bronchioles of the mock infected K18-hACE2 mouse. However, in bronchioles with epithelial hyperplasia, the hyperplastic epithelium appears ACE2 negative (arrows). Also in areas of type II respiratory epithelial metaplasia/type II pneumocyte hyperplasia (*), the cells are ACE2 negative. There is also a mild increase in ACE2 positive type II pneumocytes (arrowheads). F. Double infected K18-hACE2 mouse, 10 days post IAV infection and 7 days post SARS-CoV-2 infection. Similar to the single IAV infected animal at 10 dpi, there is ACE2 expression in the unaltered bronchiolar epithelium, while it is absent in the hyperplastic bronchiolar epithelium and in areas of respiratory epithelial metaplasia/type II pneumocyte hyperplasia (*). Positive type II pneumocytes (arrowheads) are also observed.

Article Snippet: The following primary antibodies were applied: rabbit anti-human ACE2 (Novus Biologicals; clone SN0754; NBP2-67692), goat anti-IAV (Meridian Life Sciences Inc., B65141G), rabbit anti-Iba-1 (antigen: AIF1; Wako Chemicals; macrophage marker), mAB rabbit anti-mouse CD3 (clone SP7: Spring Bioscience, Ventana Medical Systems, Tucson, USA; T cell marker) and rat anti-mouse CD45R (clone B220, BD Biosciences; B cell marker), following previously published protocols – . and rabbit anti-SARS-CoV nucleocapsid protein (Rockland, 200-402-A50).

Techniques: Expressing, Infection

Journal: bioRxiv

Article Title: Sequential infection with influenza A virus followed by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) leads to more severe disease and encephalitis in a mouse model of COVID-19

doi: 10.1101/2020.10.13.334532

Figure Lengend Snippet: ACE2 expression in the brain. A. Mock infected control animals. A1. K18-hACE2 transgenic mouse, unaltered hypothalamus. A2, A3. ACE2 expression in capillary endothelial cells in K18-hACE2 transgenic mouse (A2) and wildtype mouse (A3). B. SARS-CoV-2 infected hACE transgenic mouse. Endothelial ACE2 staining shows an intact endothelial layer (arrowheads) also in areas of perivascular infiltration (arrow). C. IAV and SARS-CoV-2 double infected hACE transgenic mouse. Endothelial ACE2 expression (arrowheads) is lost in areas of more intense infiltration (arrows). HE stain and immunohistology, hematoxylin counterstain. Bars represent 20 μm.

Article Snippet: The following primary antibodies were applied: rabbit anti-human ACE2 (Novus Biologicals; clone SN0754; NBP2-67692), goat anti-IAV (Meridian Life Sciences Inc., B65141G), rabbit anti-Iba-1 (antigen: AIF1; Wako Chemicals; macrophage marker), mAB rabbit anti-mouse CD3 (clone SP7: Spring Bioscience, Ventana Medical Systems, Tucson, USA; T cell marker) and rat anti-mouse CD45R (clone B220, BD Biosciences; B cell marker), following previously published protocols – . and rabbit anti-SARS-CoV nucleocapsid protein (Rockland, 200-402-A50).

Techniques: Expressing, Infection, Control, Transgenic Assay, Staining, H&E Stain