Fig. 5.

PPT1 downregulates Gpx1 palmitoylation and modulates angiogenesis.

(A–B) Thioesterases (APT1/2, PPT1/2, or ABHD17a) were co-expressed with Flag-Gpx1 in HEK293T cells, and protein palmitoylation was evaluated using the ABE assay (A), and quantified (B). (C–F) The Oxygen-Induced Retinopathy (OIR) disease model was established using WT and PPT1-KO mice, and mouse retinas were collected for the evaluation of protein palmitoylation by ABE (C–D) or flat-mounting to examine the relative area of neovascularization (E–F) at post-natal day 17 (P17). n = 7 for WT and 13 for PPT1-KO. (G–H) The rate of cell proliferation in HUVECs expressing either an empty vector or PPT1-Flag constructs, treated with (H) or without (G) H₂O₂ (3h), was examined using CCK8 assays. (I–L) HUVECs expressing an empty vector or PPT1-Flag, treated with (K–L) or without (I–J) H₂O₂ (3h), were subjected to the tube formation assay, and the relative number of tubule nodes was quantified. (M–P) HUVECs expressing an empty vector or PPT1-Flag, treated with (O–P) or without (M–N) H₂O₂ (3h), were subjected to the wound healing assay, and the percentage of relative wound healing was quantified. Blots and images shown are representative, and data represents the mean ± S.E.M. from three biological replicates or otherwise indicated. Statistical significance is denoted as ∗p ≤ 0.05, ∗∗p ≤ 0.01, ∗∗∗p ≤ 0.001, determined by student t-test or one-way ANOVA followed by Tukey's post hoc test.