Journal: BMC pulmonary medicine

Article Title: IL-32 induces epithelial-mesenchymal transition by triggering endoplasmic reticulum stress in A549 cells.

doi: 10.1186/s12890-020-01319-z

Figure Lengend Snippet: Fig. 4 GRP78 expression characteristics induced by IL-32 or 4-PBA. a GRP78 mRNA expression levels. *P < 0.05 compared with the control group, **P < 0.05 compared with the rhIL-32 group. b and c GRP78 protein expression levels, measured by western blotting. *P < 0.05 compared with the control group, **P < 0.05 compared with the rhIL-32 group

Article Snippet: A sample containing 20 μg of protein was separated on a 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis gel (Solarbio), transferred to a polyvinylidene fluoride membrane, and incubated with rabbit anti-mouse N-cadherin, GRP78, and α-SMA primary antibodies (Proteintech, Wuhan, China), or βactin antibody (Bioss, Beijing, China) as a loading control, at room temperature for 2 h. Subsequently, the membrane was washed, incubated with the horseradish peroxidase-conjugated goat anti-rabbit secondary antibody (Proteintech, Wuhan, China) for 1 h at room temperature, and exposed using the ECL kit.

Techniques: Expressing, Control, Western Blot

Journal: The Journal of Biological Chemistry

Article Title: Identification and characterization of Nanobodies targeting the EphA4 receptor

doi: 10.1074/jbc.M116.774141

Figure Lengend Snippet: Nb 39 and Nb 53 do not completely inhibit ephrin-A5 binding to EphA7. With Alphascreen technology we determined whether Nb 39 and Nb 53 could inhibit ephrin-A5 binding to EphA7. These Nbs did not completely inhibit the interaction between ephrin-A5 and EphA7 at the highest concentrations tested. This assay was done in triplicate, and data are represented as the mean ± S.D.

Article Snippet: To test the inhibition of EphA7 and ephrin-A5 interaction, His-tagged human ephrin-A5 (Sino Biological, Beijing, China) was biotinylated with a five-times molar excess of EZ link NHS biotin (Thermo Fisher Scientific) as is described for the Nbs.

Techniques: Binding Assay, Amplified Luminescent Proximity Homogenous Assay

Journal: Arthritis research & therapy

Article Title: Stiffness and axial pain are associated with the progression of calcification in a mouse model of diffuse idiopathic skeletal hyperostosis.

doi: 10.1186/s13075-023-03053-3

Figure Lengend Snippet: Fig. 6 ENT1−/− mice show neuroplastic changes in the cervical enlargement of the spinal cord at 6.5 months of age. A–C Representative images of immunofluorescent detection and quantification within the dorsal horn of the cervical spinal cord in wild-type and ENT1−/− mice. No differences were detected between left and right dorsal horns using Mann–Whitney’s test, so data were pooled (n = three to four female, three male per genotype). To account for anatomical and size differences in the region of interest, the upper, mid, and lower regions of the cervical enlargement were analyzed independently. A region of interest was defined around each dorsal horn to analyze laminae one to four, based on greyscale density using brightfield images. The area was standardized for each region of the cervical enlargement and the raw integrated density of immunoreactivity was averaged from up to three randomly selected sections from each region per animal. Immunoreactivity was measured for A calcitonin gene-related peptide (CGRP), B glial fibrillary acidic protein (GFAP), and C ionized calcium-binding adapter molecule 1 (IBA1). Data are plotted as mean ± SEM. *P < 0.05 by Mann–Whitney test for genotype differences and Kruskal–Wallis with Dunn’s multiple comparisons test for differences between cervical regions

Article Snippet: Immunoglobulin G isotype controls for CGRP (1:750; 5–001-A, R&D systems: Minneapolis, MN), IBA1 (1:1000; 02–6102, Thermo Fisher Scientific), and GFAP (1:500; MA110,406, Thermo Fisher Scientific), as well as secondaryonly controls, were run in parallel.

Techniques: Binding Assay, MANN-WHITNEY