Journal: iScience
Article Title: Polyethylene terephthalate microplastics impair erectile function through macrophage mediated cGAS-STING ferroptosis
doi: 10.1016/j.isci.2026.115044
Figure Lengend Snippet: PET-MPs induce EndMT via macrophage ferroptosis and activation of the cGAS-STING pathway (A) IF staining of CD31 and vimentin in CC tissue; scale bars, 50 μm. (B) Western blot analysis of E-cadherin, CD31, α-SMA, and FSP1 expression in endothelial cells cultured with PET-MP alone (MP) or PET-MP plus macrophages (MP + Mφ). (C) Transwell migration assay shows enhanced endothelial cell migration; scale bars, 50 μm. (D) Quantification of migrated cells in the Transwell assay. (E) Scratch wound-healing assay shows increased wound closure; scale bars, 100 μm. (F) Quantification of wound-healing rates. (G) Tube formation assay shows impaired angiogenesis; scale bars, 100 μm. (H) Quantification of vascular junctions in the tube-formation assay. (I) IF staining of CD31 and vimentin expression in endothelial cells; scale bars, 50 μm. (J) Western blot analysis of TGF-β, Smad2/3 phosphorylation, E-cadherin, and GPX4 expression. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001.
Article Snippet: Equal amounts of protein (30 μg) were separated on 10% SDS-PAGE gels at 120 V for 2 h and transferred onto NC membranes at 400 mA for 1 h. Membranes were blocked with 5% non-fat milk at room temperature for 2 h and incubated overnight at 4°C with primary antibodies against TGF-β (Abcam, ab92486, 1:1000), α-SMA (Abcam, ab7817, 1:5000), eNOS (Cell Signaling Technology, CST #9572, 1:1000), p-eNOS (CST #9571, 1:1000), β-tubulin (Proteintech, 10068-1-AP, 1:5000), IL-1β (CST #12703, 1:1000), IL-6 (CST #12912, 1:1000), TNF-α (Abcam, ab6671, 1:1000), PPARα (Abcam, ab8934, 1:1000), PGC-1α (Abcam, ab54481, 1:1000), SDHB (Proteintech, 10620-1-AP, 1:2000), cGAS (CST #31659, 1:1000), STING (CST #13647, 1:1000), GPX4 (Abcam, ab125066, 1:1000), E-cadherin (CST #3195, 1:1000), Smad2 (CST #5339, 1:1000), Smad3 (CST #9523, 1:1000), p-Smad2 (CST #3108, 1:1000), p-Smad3 (CST #9520, 1:1000), CD31 (Abcam, ab28364, 1:1000), and FSP1 (Abcam, ab218163, 1:1000).
Techniques: Activation Assay, Staining, Western Blot, Expressing, Cell Culture, Transwell Migration Assay, Migration, Transwell Assay, Wound Healing Assay, Tube Formation Assay, Phospho-proteomics
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