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Jeffrey B. Travers  Journal of Investigative Dermatology 

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1 Oxidative Stress Can Activate the Epidermal Platelet-Activating Factor Receptor 
Jeffrey B. Travers  Journal of Investigative Dermatology  Volume 112, Issue 3, Pages (March 1999) DOI: /j x Copyright © 1999 The Society for Investigative Dermatology, Inc Terms and Conditions

2 Figure 1 Effect of TBH on intracellular Ca2+ levels in KB cells. A clone of KB cells transduced with the MSCV 2.1 retrovirus alone (KBM; as control) or a clone of KB cells transduced with the PAF-R (KBP) were loaded with the calcium-sensitive dye indo-1AM, and changes in [Ca2+[I in response to 10 nM PAF (P), 100 nM endothelin-1 (E) or 100 μM t-BuOOH (TBH) assessed by a spectrophotometer. Journal of Investigative Dermatology  , DOI: ( /j x) Copyright © 1999 The Society for Investigative Dermatology, Inc Terms and Conditions

3 Figure 2 Concentration dependence of TBH-induced intracellular Ca2+ mobilization. KBM or KBP cells were treated with various concentrations of TBH and the peak change in [Ca2+[I assessed as described in Materials and Methods. Each point represents the mean ± SD of the peak change in [Ca2+[I from three separate experiments using representative clones; similar results were found in two other KBP and one KBM clones. *Statistically significant (p < 0.05) difference from t-BuOOH-treated KBM cells. Journal of Investigative Dermatology  , DOI: ( /j x) Copyright © 1999 The Society for Investigative Dermatology, Inc Terms and Conditions

4 Figure 3 Effect of PAF-R antagonists and antioxidants on intracellular Ca2+ mobilization in KBP cells. KBP cells were pretreated for 2 min with 100 nM CV-6209 (vertical line fill pattern), 10 μM WEB 2086 (horizontal line fill pattern) or preincubated for 30 min with 1 μM TMTU (diagonal fill pattern), 10 U per ml vitamin E (closed fill pattern), or appropriate vehicle (open fill pattern) followed by treatment with 10 nM PAF or 100 μM t-BuOOH. The peak change in [Ca2+[I in response to these agonists was then assessed. Each point represents the mean ± SD percentage of the peak change in [Ca2+[I of vehicle-treated KBP cells from at least three separate experiments using a representative clone. Similar results were found in a second KBP clone tested. *Statistically significant (p < 0.05) difference from vehicle-treated KBP cells. Journal of Investigative Dermatology  , DOI: ( /j x) Copyright © 1999 The Society for Investigative Dermatology, Inc Terms and Conditions

5 Figure 4 Effect of t-BuOOH on sn-2 acetyl GPC biosynthesis in KB cells. KBM or KBP cells were treated with 100 μM t-BuOOH for various times and (A) 1-hexadecyl-2-acetyl GPC (PAF) and (B) 1-palmitoyl-2-acetyl GPC measured by mass spectrometry. Each point represents the mean ± SD amounts of PAF or PAPC from three separate experiments using representative clones; similar results were found in at least one other KBP and KBM clone. *Statistically significant (p < 0.05) difference in PAF or PAPC production from unstimulated KB cells. Journal of Investigative Dermatology  , DOI: ( /j x) Copyright © 1999 The Society for Investigative Dermatology, Inc Terms and Conditions

6 Figure 5 Hypothetical involvement of the PAF system in keratinocyte oxidative stress. Journal of Investigative Dermatology  , DOI: ( /j x) Copyright © 1999 The Society for Investigative Dermatology, Inc Terms and Conditions


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