Journal: Journal of Neuroinflammation

Article Title: Pathological high intraocular pressure induces glial cell reactive proliferation contributing to neuroinflammation of the blood-retinal barrier via the NOX2/ET-1 axis-controlled ERK1/2 pathway

doi: 10.1186/s12974-024-03075-x

Figure Lengend Snippet: Antibodies used for histological analyses

Article Snippet: The gel electrophoresis was run using NuPAGE running buffer MES at room temperature with a voltage of 130 V for 60 min. After electrophoresis, the proteins were transferred to polyvinylidene fluoride membranes (Bio-Rad, Gladesville, Australia) using a dry transfer system (Bio-Rad, Hercules, CA, USA), and a standard transfer buffer (with 20% methanol), a voltage of 20 V was applied for 7 min. For immunoblotting, membranes were incubated with the appropriate antisera ET-1 (1:1000, NB300-526, Novus Biologicals, Littleton, USA), p47 phox (1:1000, sc-17,845, Santa Cruz Biotechnology), gp91 phox (1:1000, sc-130,543, Santa Cruz Biotechnology), polyclonal antibodies recognizing phospho-JNK (Thr183/185) (1:1000; catalog no. 9251), phospho-ERK1/2 (Thr202/204) (1:1000; catalog no. 9101) and phospho-p38 MAPK (Thr180/182) (1:1000; catalog no. 4511) overnight at 4 °C, and labeling was carried out using a multi-step detection procedure.

Techniques: