Journal: Frontiers in Ophthalmology
Article Title: Brief research report: ETS-1 blockade increases ICAM-1 expression in activated human retinal endothelial cells
doi: 10.3389/fopht.2024.1384428
Figure Lengend Snippet: Impact of ETS-1 silencing on the levels of human retinal endothelial cell ICAM1 transcript and membrane-bound ICAM-1 protein. Human retinal endothelial cells were transfected with ETS-1 siRNA s4847, s4848, s4847 plus s4848, or non-targeted control siRNA for 24 hours, and subsequently stimulated for another 24 hours with TNF-α (10 ng/mL), IL-1β (10 ng/mL) or fresh medium alone. (A-C) Graphs show expression of ICAM1 transcript after transfection with (A) ETS-1 siRNA s4847, (B) ETS-1 siRNA s4848, and (C) ETS-1 siRNA s4847 plus s4848, versus the common non-targeted control siRNA. Bars represent mean expression normalized to the expression of reference genes, RPLP0 and GAPDH (n = 3-4 monolayers/condition). Error bars show standard deviation. (D, E) Graph shows membrane-bound ICAM-1 protein after treatment with siRNA, followed by (D) TNF-α or fresh medium, and (E) IL-1β or fresh medium. Bars represent mean relative fluorescence of immunolabelled cell monolayers after corrections for background fluorescence and cell number (n = 4 cell monolayers/condition). Error bars show standard deviation. x = below detectable level. Data were analyzed by (A-C) two-tailed unpaired t-test and (D, E) two-way ANOVA, with Sídák post-hoc testing: *p < 0.05; **p<0.01; ***p < 0.001; ****p < 0.0001.
Article Snippet: Mouse monoclonal IgG2b K anti-human ICAM-1 (clone LB-2) and isotype-matched negative control anti-dansyl (clone 27-35) antibodies were bought from BD Pharmingen (San Jose, CA), and used at a working concentration of 1 μg/mL.
Techniques: Membrane, Transfection, Control, Expressing, Standard Deviation, Fluorescence, Two Tailed Test