Journal: Particle and Fibre Toxicology
Article Title: Carbon nanoparticles induce ceramide- and lipid raft-dependent signalling in lung epithelial cells: a target for a preventive strategy against environmentally-induced lung inflammation
Figure Lengend Snippet: Ectoine as well as antioxidants prevent ceramide generation and subsequent receptor translocation. ( A ) Flow cytometric analysis of intracellular ROS generation in RLE-6TN cells measured by relative intracellular DCF fluorescence after treatment with the indicated doses of CNP [μg/cm 2 , 1 h], CP [μg/cm 2 , 1 h], BSA-coated CNP, or C6 ceramide [5 μM, 15 min] and pre-treatment with the respective inhibitors tocopherol [Toco; 50 μM] N-acetylcysteine [NAC; 0.1 mM], or ectoine [E; 1 mM]. C indicates respective vehicle controls. ( B ) Amount of ceramide generation was detected by lipid extraction and HPTLC in RLE-6TN cells. ( C ) EGFR translocation as described in Figure 2 after treatment and pre-treatment as described above. ( D ) EGFR activation as described in Figure 1 after treatment and pre-treatment as described above. Means in A , C , and D are given as values relative to the respective controls. *, Significantly different from control; †, significantly different from 10 μg/cm 2 CNP alone; §, significantly different from C6 alone.
Article Snippet: Inhibitors were added to the cells 18 h (NAC), 4 h (ectoine), or 60 min (compound 32, alpha-tocopherol) prior to treatment with CNP or C6 ceramide (N-hexanoyl-D-erythro-shingosine, Calbiochem, Schwalbach, Germany).
Techniques: Translocation Assay, Flow Cytometry, Fluorescence, High Performance Thin Layer Chromatography, Activation Assay