Fig 2: AMPs are increased in keratinocytes exposed to ozone in vitro. qRT-PCR analysis of transcript levels of hBD1 (a), hBD2 (b), hBD3 (c), and CAMP (d) in keratinocytes exposed to 0.2 ppm of O3 directly after exposure (T0), 6 h (T6), 12 h (T12), and 24 h (T24) post-exposure. (e) Representative immunoblot for hCAP18 levels in keratinocytes at different time points (T0, T6, T12, and T24), quantification is depicted in panel (f). Results from three independent experiments and three replicates. * = p-value<0.05. Error bars are defined as mean±SEM.

Fig 3: Exposure of human biopsies to ozone increases AMP protein levels in the skin. Ex vivo human abdominoplasty biopsies from three different donors were exposed to 0.4 ppm O3.Sections were collected directly after exposure (T0) and 24 h (T24) post exposure and embedded in paraffin. Immunofluorescence was performed analyzing protein levels of AMPs hCAP18/LL37 (A) hBD2 (B) and hBD3 (C). Images quantification was performed using ImageJ. Graphs depicted are the results of the quantification of three independent experiments. The images reported in this figure were chosen as being most representative of the quantification. Red staining indicates either hCAP18/LL37 or hBD3, green staining indicates hBD2. Blue staining represents DAPI. * = p-value<0.05. Error bars are defined as mean±SEM. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Fig 4: Exposure of human volunteer subjects to ozone increases AMP protein levels in the skin. Forearms of human volunteer subjects were exposed to 0.8 ppm O3 for 3 h a day for 5 consecutive days. We examined protein levels of hCAP18/LL37 (a, c) and hBD3 (b, d) using immunofluorescence in 3 mm punch biopsies taken from the aforementioned subjects from both unexposed (control) and exposed lateral forearms. Images were quantified using ImageJ. Data shown in (c–d) are the results of averages of multiple images from at least three different subjects. Images depicted in Fig. 5 were chosen as being the most representative of results after quantification. * = p-value<0.05. Error bars are defined as mean±SEM.