Figure S4.

TLNRD1 modulates endothelial barrier function. (A and B) siCTRL and siTLNRD1 endothelial cells were allowed to form a monolayer without flow stimulation. Cells were then fixed and stained for phospho-Myosin light chain (pMLC S20) before being imaged on a spinning disk confocal microscope. (A) Representative sum projections are displayed. (B) The overall integrated density was measured for each field of view from SUM projections (three biological repeats, n = 45 FOV per condition). (C and D) Assessment of trans-endothelial electrical resistance (TEER) in siCTRL and siTLNRD1 endothelial monolayers before and after thrombin stimulation was conducted utilizing the xCELLigence system. Individual TEER trajectories were normalized to the readings before the thrombin stimulation to study the effect of thrombin on TEER over time. Thrombin stimulation was performed 48 h after initial recording. (C) Displays representative data from one biological replicate. Here, the mean TEER trajectory from three individual wells is delineated with a bold line. In contrast, individual TEER curves are rendered in a lighter shade to delineate specific measurements within the same replicate. (D) Comparative analysis of the TEER values at 26 h after thrombin stimulation (two biological repeats, six measurements). The P values were determined using a t test (two-sided, assuming unequal population variances).