Skip to main content
. Author manuscript; available in PMC: 2020 Aug 15.
Published in final edited form as: Cell Chem Biol. 2019 May 23;26(8):1081–1094.e6. doi: 10.1016/j.chembiol.2019.04.007

Figure 6: The role of SRC activation in regulation of endothelial barrier mediated by physiological stimuli.

Figure 6:

(A) Activation of SFKs following S1P stimulation. HPAE cells were serum starved (1 hour) and treated with 1 μM S1P. Cell lysates were collected at the indicated time points and analyzed with the indicated antibody. (A and C) Average relative phosphorylation was calculated from 4 independent experiments. Significance was calculated using a one-way ANOVA with a Dunnett’s multiple comparisons correction, *p < 0.05, **p < 0.01; error bars depict standard deviation. (B) The role of SFK activity in S1P-mediated barrier enhancement. HPAE cells were serum starved (1 hour) and analyzed using TER; 1 μM S1P and 100 nM Saracatinib or vehicle (DMSO) were added at time point 0. (C) Activation of SFKs following stimulation with thrombin. HPAE cells were serum starved (1 hour) and treated with 50 nM of Thrombin. Cell lysates were collected at the indicated time and analyzed using indicated antibody. (D) The role of SFK activity in barrier recovery following thrombin treatment. HPAE cells were serum starved (1 hour) and analyzed using TER. Thrombin (50 nM) was added at time point 0 and Saracatinib (100 nM, S) or vehicle (ethanol, V) was added 15 minutes later. (E and F) Effect of SRC and LYN activation on barrier recovery. HPAE cells co-expressing RapR-SRC-cerulean and mCherry-FRB (E) or RapR-LYN-cerulean and mCherry-FRB (F) were analyzed using TER. Thrombin (50 nM) was added at time 0 and rapamycin (500 nM) or vehicle (ethanol) (green line) was added 15 minutes later. All exogenous proteins were expressed using adenoviral transduction. (B, D-F) Graphs show an average resistance from 3 experiments and error bars depict the 90% confidence intervals. See also figures S6A–D.