Figure 1

Schematic representation of the experimental design. The cells from one donor (#MD058501) were used for the RNA-seq experiment (A) and exposed to air, 3R4F smoke 1/30, e-cigarette aerosol 1/7 and 1/3. At the end of the exposure runs, nicotine was quantified from the media (1 media sample per chamber and run) and from 3 inserts filled with PBS for each treatment (A). CBF was also measured in the cell inserts immediately after exposure (A). The other endpoints (TEER, LDH) and RNA for RNA-seq were collected at 24 and 48 hrs post-exposure (A). For the RNA-seq experiment, three exposure runs were performed independently (Run1, 2, and 3) with 3 inserts for each treatment and time point. *Indicates that the immunochemistry were performed in a separate exposure experiment with donor #MD058501. Three independent runs with three inserts per treatment were also used. To validate the RNA-seq findings, a qPCR validation experiment was conducted with three donors (#MD058501, #MD008301, #MD009101) using the same exposure conditions and sets of 20 genes (B). The RNA was collected 24 hrs post-exposure (B). One exposure run was performed with 3 inserts per treatment. (C) Schematic of an exposure chambers with insert positions.