Fig. 4
From: Two distinct cellular pathways leading to endothelial cell cytotoxicity by silica nanoparticle size
20-nm SiNP-induced autophagy depends on PI3K/AKT/eNOS/nitric oxide signaling pathway. a Western blot analysis of LC3B-I to LC3B-II conversion in HUVECs treated with 10 μM of Apocynin and Mito-Tempo for 30 min and subsequently treated with 20-nm SiNPs at 20 μg/mL for 5 h. Relative LC3-II to LC3-I ratios are indicated in the graph using Image J software. SiNPs-induced autophagy depends on PI3K/AKT or eNOS signaling pathway activation, but independent of ROS or AMPK. b Western blot analysis of LC3B-I to LC3B-II conversion and and AKT, AMP-activated protein kinase (AMPK), endothelial NO-synthase (eNOS), and p38 phosphorylation. HUVECs were treated with 20- and 50-nm of SiNPs for the indicated times at 20 μg/mL. c, d Western blot analysis of LC3-I to LC3-II conversion and AMPK phosphorylation or p38 phosphorylation in HUVECs treated with indicated concentrations of Compound-C and SB203580 for 30 min and subsequently treated with 20-nm SiNPs at 20 μg/mL for 5 h, respectively. e–g Western blot analysis of the LC3-I to LC3-II conversion following treatment with PI3K inhibitor (Wortmannin), eNOS inhibitor (L-LAME), and eNOS siRNA, respectively. Relative LC3-II to LC3-I ratios are quantified by Image J software