cell culture 149 cultures  (ATCC)

Journal: American Journal of Respiratory Cell and Molecular Biology

Article Title: Reactive Oxygen Species/Hypoxia-Inducible Factor-1α/Platelet-Derived Growth Factor-BB Autocrine Loop Contributes to Cocaine-Mediated Alveolar Epithelial Barrier Damage

doi: 10.1165/rcmb.2016-0096OC

Figure Lengend Snippet: Cocaine-mediated permeability change in pulmonary epithelial cells. (A) Cocaine increased the permeability of alveolar epithelial cell line L2 cells in a dose-dependent manner, with maximal response at 100 μM. (B) Electronic cell-substrate impedance sensing (ECIS) assay analysis confirmed cocaine-mediated increase of barrier permeability in a dose-dependent manner in mouse primary alveolar epithelial cells. (C) Exposure of L2 cells to cocaine resulted in a time-dependent decrease in expression of zona occuldens proteins (Zo)-1 but not occludin or claudin-5. The analysis data are shown at the bottom. (D) Primary epithelial cells grown in the ECIS chamber were treated with cocaine for 48 hours and stained with antibodies specific for Zo-1. Arrows indicate cocaine treatment significantly reduced the Zo-1 expression compared with control. All the data are presented as mean ± SD of at least three individual experiments. *P < 0.05, **P < 0.01 versus control group. Ctrl, control; FITC, fluorescein isothiocyanate; TJPs, tight junction proteins.

Article Snippet: Cell Culture Primary lung alveolar epithelial cell line L2 cells were purchased from ATCC (Manassas, VA).

Techniques: Permeability, Expressing, Staining, Control

Journal: American Journal of Respiratory Cell and Molecular Biology

Article Title: Reactive Oxygen Species/Hypoxia-Inducible Factor-1α/Platelet-Derived Growth Factor-BB Autocrine Loop Contributes to Cocaine-Mediated Alveolar Epithelial Barrier Damage

doi: 10.1165/rcmb.2016-0096OC

Figure Lengend Snippet: The ROS/HIF-1α/PDGF autocrine loop involved in cocaine-mediated alveolar epithelial barrier damage. (A) PDGF-BB–induced, dose-dependent generation of ROS using DCF-DA assay. (B) Representative Western blotting of PDGF-BB–mediated, dose-dependent down-regulation of Zo-1. (C) Representative RT-PCR image of PDGF receptor (PDGFR)-α and PDGF-β expression in L2 cells. (D) Pretreatment of cells with the PDGFR blocker STI-571 resulted in amelioration of cocaine-mediated induction of ROS production. (E) Western blot analysis showing that STI-571 inhibited cocaine-induced up-regulation of PDGF-BB and HIF-1α and suppressed cocaine-mediated down-regulation of Zo-1. (F) Pretreatment of cells with STI-571 resulted in amelioration of cocaine-mediated permeability change. All the data are presented as mean ± SD of at least three individual experiments. *P < 0.05, **P < 0.01 versus control group; #P < 0.05, ##P < 0.01 versus cocaine-treated group.

Article Snippet: Cell Culture Primary lung alveolar epithelial cell line L2 cells were purchased from ATCC (Manassas, VA).

Techniques: Western Blot, Reverse Transcription Polymerase Chain Reaction, Expressing, Permeability, Control