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. 2016 Feb;186(2):248–258. doi: 10.1016/j.ajpath.2015.10.001

Figure 2.

Figure 2

Trace, total length, and vector length of lysosomal exocytosis in iHUVECs and DHE fluorescence intensity. A: Representative and normalized XY-position traces of labeled lysosomes tracking over 60 minutes. The trajectories of peripheral lysosomes show significantly longer tracks in LPS, LPS + NOHA, and EPC-CM10 and EPC-CM50 groups. B and C: Individual organelles were randomly selected from a time-lapse series obtained before and after application of LPS, NOHA, and EPC-CM. In control, movements were non-vectorial. After LPS, lysosomal movement became more dynamic and directional. When nitric oxide donor NOHA or EPC-CM was used for pretreatment before LPS, lysosomal movements were reduced. D: DHE fluorescence intensity after LPS treatment of cultured HUVECs. Data are expressed as means ± SEM. n = 4 to 5 per group and 30 lysosomal particles per cell (B and C); n = 12 for 10 and 2.5 μg/mL groups (D); n = 6 for other tested groups (D). *P < 0.05 versus total trajectory (B and C); †P < 0.05 versus control (B and C); and ‡P < 0.05 versus LPS + NOHA and EPC-CM10 and EPC-CM50 (B and C), differences tested by one-way analysis of variance followed by the Holm-Sidak multiple comparison test. *P < 0.05, **P < 0.01 versus all lower LPS treatment group(s) at indicated times within each time point (D); †P < 0.05 versus no LPS (D). CM, conditioned medium; DHE, dihydroethidium; EPC, endothelial progenitor cell; iHUVEC, immortalized human umbilical vein endothelial cell; LPS, lipopolysaccharide; NOHA, NG-hydroxy-l-arginine.