Journal: Investigative Ophthalmology & Visual Science
Article Title: Integrated Transcriptomics and Experimental Validation Reveal That Ellagic Acid Alleviates Fuchs Endothelial Corneal Dystrophy via PLAU/NF-κB Signaling
doi: 10.1167/iovs.67.1.31
Figure Lengend Snippet: In vitro experiments confirming ellagic acid (EA) target on PLAU to regulate NF-κB signaling pathway in corneal endothelial cells. ( A – C ) Western blot analysis of PLAU, NF-κB p65, and phospho-NF-κB p65 in H 2 O 2 -treated B4G12 cells, with or without EA treatment. The relative protein expression levels are shown for B PLAU and C the ratio of phospho-NF-κB p65 to NF-κB p65 ( n = 3 for PLAU, and n = 4 for phospho-NF-κB p65 to NF-κB p65). ( D – F ) Western blot analysis of PLAU, NF-κB p65, and phospho-NF-κB p65 in UVA-irradiated B4G12 cells, with or without EA treatment. Relative protein expression levels are shown for E PLAU and F the ratio of phospho-NF-κB p65 to total NF-κB p65 ( n = 3). ( G – I ) Western blot analysis of PLAU, NF-κB p65, and phospho-NF-κB p65 following siPLAU treatment in H 2 O 2 -treated B4G12 cells, with or without EA. Relative protein expression levels are presented for G PLAU and I the ratio of phospho-NF-κB p65 to NF-κB p65 ( n = 3 for PLAU, and n = 4 for phospho-NF-κB p65 to NF-κB p65). ( J ) Schematic diagram illustrating the proposed anti-inflammatory and anti-oxidant mechanisms of EA in FECD, highlighting its role in modulating the NF-κB pathway. All data are presented as mean ± SD. * P < 0.05, ** P < 0.01 by 1-way ANOVA with Tukey’s post hoc test.
Article Snippet: Primary antibodies against PLAU (Proteintech, China; #17968-1-AP), NF-κB p65 (Abcam, UK; #ab32536), phospho-NF-κB p65 (S536; Abcam, UK; #ab76302), GAPDH (ABclonal, China; #AC002), and β-Actin (ABclonal, China; #AC004) were applied overnight at 4°C.
Techniques: In Vitro, Western Blot, Expressing, Irradiation