IFN--pretreated HUVECs were exposed to PRA at the indicated times prior to performing Western blotting (a, g, j) and Rab5 pulldowns (b, e, h). HUVEC stably transduced with Rab5 WT-GFP were treated with PRA for 30 min and subjected to proximity ligation assays (PLAs, c). HUVECs were co-transduced with Rubicon-GFP or RNF34-GFP along with Rab5-RFP constructs, treated with PRA for 30 min and analyzed by confocal I.F. (d, f, i). Two-tailed Student’s t -test (c, f, i) was used for statistical comparisons where p < 0.05 is considered statistically significant. Experiments repeated 2 times (c, f, i), 3 times (b, d, e, g, h, j) , and 5 times (a) with different HUVEC donors. All scale bars, 200 m. p -values are indicated in the figures.
