Journal: Antioxidants
Article Title: Oxidative Stress Increases Endogenous Complement-Dependent Inflammatory and Angiogenic Responses in Retinal Pigment Epithelial Cells Independently of Exogenous Complement Sources
doi: 10.3390/antiox8110548
Figure Lengend Snippet: Oxidative stress induced complement component accumulation in ARPE-19 cells. ( A ) Properdin mRNA levels were increased 24 h following H 2 O 2 treatment. This did not affect ( B ) apical properdin secretion, but was confirmed in the protein level by immunohistochemistry using an ( C – E ) anti-properdin (red) antibody. ( F ) C3 mRNA and ( G ) apical C3 protein secretion were not altered in stressed ARPE-19 cells. Immunohistochemistry using ( H – J ) anti-C3 (green) antibodies showed an increase of cell-associated ( I , J ) C3 after oxidative stress treatment. ( K ) Cfh mRNA and ( L ) CFH apical protein concentration were decreased following H 2 O 2 treatment. ( M – O ) Immunohistochemistry using anti-complement factor H (CFH, purple) antibodies showed an increase in cell-associated ( N , O ) CFH after oxidative stress treatment. Mean with standard deviation is shown, ** p ≤ 0.01, **** p ≤ 0.0001; dotted line depicts untreated control ( A , F , K ); w/o untreated control ( G , G , L ); ELISA control standard curves and protein concentrations in the basal supernatants are shown in the .
Article Snippet: C3 quantification was performed using a newly developed C3 ELISA (cat# HK366-01; Hycult Biotech, Uden, Netherlands) according to the manufacturer’s protocol.
Techniques: Immunohistochemistry, Protein Concentration, Standard Deviation, Enzyme-linked Immunosorbent Assay