Journal: Antioxidants

Article Title: In Vitro Effects of Lipopolysaccharide on Rabbit Sperm: Toll-like Receptor 4 Expression, Motility, and Oxidative Status

doi: 10.3390/antiox14040431

Figure Lengend Snippet: UV (ultraviolet) micrographs of rabbit spermatozoa without ( A ) and after 4 h of incubation with 400 μg/mL LPS (lipopolysaccharide) ( B ) treated with anti-TLR4 antibody. The label is localized in the middle piece of the sperm tail. In ( A ), the TLR4 (Toll-like receptor 4) signal was weak, whereas in ( B ), after LPS incubation, an intense TLR4 signal was observed. Nuclei were stained with DAPI (4′,6-diamidino-2-phenylindole). Scale bars: 5 μm.

Article Snippet: Then, the slides were incubated overnight at 4 °C in a humid chamber with the primary antibodies: CD284 (TLR4) Monoclonal Antibody (HTA125) eBioscence, (Invitrogen, Thermo Fisher Scientific, Carlsbad, CA, USA) diluted 1:200.

Techniques: Incubation, Staining

Journal: BMC Pulmonary Medicine

Article Title: By activating endothelium histone H4 mediates oleic acid-induced acute respiratory distress syndrome

doi: 10.1186/s12890-024-03334-w

Figure Lengend Snippet: TLR and calcium signaling involved in histone H4-mediated endothelium activation. After the MLVECs were challenged with histone H4 (15 mg/L) for 12 h, the changes in endothelial HS were evaluated by flow cytometry ( A ) while the VE-Cadherin in the cell membrane was measured by western blot ( B ). The vWF in the supernatant ( C, E ) and P-selectin translocation ( D, F ) were measured through ELISA. Prior to histone H4 challenge, the cells were pretreated for one hour with a blocking antibody (10 mg/L) against TLRs (TLR1, TLR2, TLR4, or TLR6) and the calcium chelator EGTA-AM (25, 50 or 100 µM). Data are presented as mean ± SD ( n = 6). The results for flow-cytometry and western blot are representative of three similar samples. * p < 0.05, ** p < 0.01 compared to the control group; # p < 0.05, ## p < 0.01 compared to the H4 group

Article Snippet: The reagents used in this study included: oleic acid (OA) and Histopaque (Sigma-Aldrich St. Louis, MO, USA); histone H4 (Millipore, Billerica, MA, USA); antibodies for P-selectin, sodium-potassium ATPase, CD31 (PECAM-1), and Ly6G (Abcam, Cambridge, MA, USA); an antibody for Cadherin-5 (BD Transduction Laboratories, CA, USA); an antibody for heparan sulfate (HS) (Bioss, Woburn, MA, USA); blocking antibodies against P-selectin, TLR4 (HTA125), TLR2 (TL2.1), and TLR1 (GD2.F4) (eBioscience, San Diego, CA, USA); a blocking antibody against TLR6 (TLR6.127) (Abcam, Cambridge, MA, USA); enzyme-linked immunosorbent assay (ELISA) kits for histone H4 and von Willebrand factor (vWF) (Cusabio Biotech, Wuhan, China); and 1,2-bis(2-aminophenoxy) ethane N, N,N’,N’-tetraacetic acid acetoxymethyl ester (EGTA-AM) (MedChemExpress, Monmouth Junction, NJ, USA).

Techniques: Activation Assay, Flow Cytometry, Membrane, Western Blot, Translocation Assay, Enzyme-linked Immunosorbent Assay, Blocking Assay, Control