Journal: Veterinary Research

Article Title: Differences in the tissue tropism to chicken oviduct epithelial cells between avian coronavirus IBV strains QX and B1648 are not related to the sialic acid binding properties of their spike proteins

doi: 10.1186/1297-9716-45-67

Figure Lengend Snippet: Lectin binding on oviduct explants. Lectin staining of oviduct explants with Sambucus nigra agglutinin (SNA) to detect the expression of alpha 2,6-linked sialic acids and Maackia amurensis agglutinin (MAA) in its two isoforms MAA I and MAA II to detect the expression of alpha 2,3-linked sialic acids. Results are summarized in Table .

Article Snippet: To detect alpha 2,3-linked sialic acids we used the lectins Maackia amurensis agglutinin (MAA) in its two isoforms MAA I and MAA II (Vector Laboratories, USA).

Techniques: Binding Assay, Staining, Expressing

Journal: Stem Cells International

Article Title: Coupled CRC 2D and ALI 3D Cultures Express Receptors of Emerging Viruses and Are More Suitable for the Study of Viral Infections Compared to Conventional Cell Lines

doi: 10.1155/2020/2421689

Figure Lengend Snippet: Influenza virus sialic acid receptor detection on 2D and 3D ALI cultures of HNTEC. (a) Influenza virus sialic acid receptor detection on 2D cultures of HNTEC. HNTEC were seeded in 12-well plates cultured at an appropriate density, then fixed and permeabilized as described above, and labeled with the biotinylated lectins SNA, MAA-I, and MAA-II and PBS (as a blank control), respectively. These three lectin markers were measured by immunofluorescence protocols. The cellular nuclei were stained with DAPI. The lectins were stained with fluorescein avidin DCS. Magnification 40x. Scale bar, 50 μ m. (b) Influenza virus sialic acid receptor detection on 3D ALI cultures of HNTEC. ALI 3D cultures of HNTEC were fixed, permeabilized, and labeled with the biotinylated lectins SNA, MAA-I, and MAA-II, respectively. The nuclei were stained with DAPI. The lectins were stained with fluorescein avidin DCS. Magnification 40x. Scale bar, 50 μ m. (c) Expression of ACE2 mRNA in HNTEC and HNBEC. Total cellular RNAs were extracted, and real-time RT-PCR was performed to detect the expression of ACE2 mRNA. The cancer cell line A549 was used as control cells.

Article Snippet: Cells in 2D or 3D cultures were fixed and permeabilized as described above, then labeled with the biotinylated lectins (20 μ g/ml, SNA, Vector Laboratories, Burlingame, CA, B-1305; 20 μ g/ml, MAA-I, Vector Laboratories, Burlingame, CA, B-1265; and 20 μ g/ml, MAA-II, Vector Laboratories, Burlingame, CA, B-1315), and then detected with fluorescein avidin DCS (A-2011, Vector Laboratories, Burlingame, CA) at 37°C for an hour in the dark.

Techniques: Cell Culture, Labeling, Immunofluorescence, Staining, Avidin-Biotin Assay, Expressing, Quantitative RT-PCR