O₃ induced NRF2 activation in human keratinocytes and MIX 1 and MIX 2 pre-treatment for 24 h potentiated this effect.

<p>Immuno-cytochemistry of keratinocytes showing localization of NRF2 (red) and Keap1 after O₃ exposure for 1 h. Images are merged and representative of at least 100 cells viewed in each experiments (n = 5). Nuclei (blue) were stained with DAPI. Original magnification X 630. Immunoreactivity of NRF2 and Keap1 was semi-quantified as area of both signals into nucleus respect to cytoplasm, by using Image J sophtware. Data are expressed in arbitrary units (averages of five experiments ± SEM, *<i>p</i> < 0.05 vs C (nuclear expression); °<i>p</i> < 0.05 vs C (cytoplasm expression)).</p

MIX 1 and MIX 2 pre-treatment prevents the decrease in cell proliferation 24 hr after O₃ exposure.

<p>Data are expressed as percentage of control (averages of five experiments ± SEM, *<i>p</i> < 0.05 vs control; #<i>p</i> < 0.05 vs O₃.</p

Cytotoxicity measured by using LDH release at T0 (A) and T24 (B) in human keratinocytes exposed to O₃ pre-treated with/without MIXs.

<p>Triton X represents 100% of LDH release. Data are expressed as percentage of Triton X-100 (averages of five experiments ± SEM, *<i>p</i> < 0.05 vs control; #<i>p</i> < 0.05 vs O3).</p

O₃ induced ROS formation in human keratinocytes and MIX 1 and MIX 2 pre-treatment prevented this effect.

<p>ROS production was measured by fluorimetry with DCFH-DA staining. Data are expressed in RFU (averages of five experiments ± SEM, *<i>p</i> < 0.05 vs control; #<i>p</i> < 0.05 vs O3).</p

Primer sequences and PCR condition.

<p>Data calculated by Bio-Rad CFX Manager Software (Bio-Rad).</p><p>Primer sequences and PCR condition.</p

O₃-induced an increase in IL-8 gene expression in human keratinocytes while MIX 1 and MIX 2 pre-treatment prevented IL-8 induction by O₃.

<p>The data are averages of five different experiments (means ± DS). * p<0.05 vs control; # vs O₃.</p

O₃ induced NF-kB (p65 subunit) activation in human keratinocytes and MIX 1 and MIX 2 pre-treatment for 24 h reverted this effect.

<p>Immuno-cytochemistry of keratinocytes showing localization of p65 (red) after O₃ exposure for 1 h. Images are merged and representative of at least 100 cells viewed in each experiments (n = 5). Nuclei (blue) were stained with DAPI. Original magnification X 630. Immunoreactivity of p65 was semi-quantified as area of activated p65 signal into nucleous respect to that into cytoplasm, by using Image J sophtware. Data are expressed in arbitrary units (averages of five experiments ± SEM, *<i>p</i> < 0.05 vs C (nuclear expression); °<i>p</i> < 0.05 vs C (cytoplasm expression)).</p

SREBP-1, SREBP-2, HMGR, and LDLr protein levels in Rett- and healthy donor-derived fibroblasts.

<p>The figure shows a representative Western blot and the densitometric analysis of SREBP-1 (a), SREBP-2 (b), HMGR (c), and LDLr (d) protein levels. The protein levels were normalized with tubulin content. The data are expressed as arbitrary units obtained analyzing the bands by using the software ImageJ. Data are the mean ± S.D. of n = 3 independent experiments carried out on 15 subject for each experimental group. Data were analyzed with unpaired Student's t test. * = p<0.05 *** = p<0.001.</p

Plasma PCSK9 protein levels in Rett patients and healthy donors.

<p>The figure shows a representative Western blot and the densitometric analysis of PCSK9 protein levels. The samples were normalized for protein loading by using Ponceau S staining. The data are expressed as arbitrary units obtained analyzing the bands by using the software ImageJ. Data are the mean ± S.D. of n = 3 independent experiments carried out on 15 subject for each experimental group. Data were analyzed with unpaired Student's t test. *** = p<0.001.</p

Plasma lipid profile in Rett and healthy donors.

<p>The figures shows the levels of total cholesterol (T-chol), triglycerides (Try), high density lipoproteins (HDL), and low density lipoproteins (LDL) in Rett patients and in healthy donors. Data are the mean ± S.D. of n = 3 independent experiments carried out on 15 subject for each experimental group. Data were analyzed with unpaired Student's t test. * = p<0.05.</p