FN, PAI-1 and CTGF expression analysis.

<p>Quantitative realtime PCR expression analysis of FN, PAI-1 and CTGF mRNAs in controls, ω-6 and ω-3 fatty acids pre-treated hTM normalized to controls (<b>A</b>) before and (<b>B</b>) after H₂O₂ exposition. (<b>C</b>) Western blot detection of cellular FN, PAI-1, CTGF, and actin in controls, ω-6 and ω-3 fatty acids pre-treated hTM. Plots of densitometric quantifications to deduce fold expressions of intracellular (<b>D</b>) FN, (<b>E</b>) PAI-1 and (<b>F</b>) CTGF, before and after H₂O₂ exposition. (<b>G</b>) ELISA quantification of FN medium contents normalized to controls. Values represent m.a. folds ± sd of three independent experiments performed on cells of three different donors (n = 9); asterisks: <i>p</i>-values of statistical significances (*<i>p</i>≤0.05; **<i>p</i>≤0.01; ***<i>p</i>≤0.001).</p

CCK-8 assay.

<p>Quantification of mitochondrial activity in hTM after 48 hours normalized to starting activity in controls. (<b>A</b>) Effects of ω-6 (16 µM) and ω-3 (50 µM) fatty acids compared to controls (Co). (<b>B</b>) Effects of H₂O₂ in controls, ω-6 and ω-3 fatty acids pre-treated hTM. (<b>C</b>) %-reduction of mitochondrial activity after H₂O₂ exposition. Values represent mean averages (m.a.) ± standard deviations (sd) of three independent experiments performed in triplicates of 5 different donors (n = 45); asterisks: p-values of statistical significances (**p≤0.01; ***p≤0.001).</p

Interleukin 1α, -6 and -8 expression analysis.

<p>Quantitative realtime PCR expression analysis of interleukins (<b>A</b>) -1α, (<b>B</b>) -6 and (<b>C</b>) -8 mRNAs in controls, hTM pre-treated with ω-6 and ω-3 before and after H₂O₂ exposition. ELISA quantification of (<b>D</b>) IL-6 and (<b>E</b>) IL-8 medium contents. Values are normalized to untreated controls and represent m.a. folds ± sd of three independent experiments performed on cells of three different donors (n = 9); asterisks: <i>p</i>-values of statistical significances (*<i>p</i>≤0.05; **<i>p</i>≤0.01; ***<i>p</i>≤0.001).</p

Primers used for realtime PCR.

<p>Primers used for realtime PCR.</p

Hsp27 and Hsp90 expression analysis.

<p>(<b>A</b>) Quantification of realtime PCR expression analysis of Hsp27 and Hsp90 mRNAs in controls, ω-6 and ω-3 fatty acids pre-treated hTM normalized to controls. (<b>B</b>) Western blot detection of cellular Hsp27, Hsp90 and actin protein in controls, ω-6 and ω-3 fatty acids pre-treated hTM. (<b>C</b>) Plot of densitometric quantifications of Hsp27 and Hsp90 protein expression in controls, ω-6 and ω-3 fatty acids pre-treated hTM adjusted to actin expression and normalized to controls. Values represent m.a. ± sd of three independent experiments performed on cells of three different donors (n = 9); asterisks: p-values of statistical significances (*p≤0.05; **p≤0.01; ***p≤0.001).</p

F-actin labelling.

<p>Phalloidin labeling of the F-actin cytoskeleton in controls, ω-6 and ω-3 supplemented hTM (<b>A</b>) before and (<b>B</b>) after H₂O₂ exposition; scale bar: 100 µm.</p

Analysis of nuclear NFκB.

<p>(<b>A</b>) Quantitative realtime PCR expression analysis of NFκB mRNA in controls, hTM pre-treated with ω-6 and ω-3 before and after H₂O₂ exposition. (<b>B</b>) Quantification of nuclear NFκB protein contents. Values are normalized to untreated controls and represent m.a. folds ± sd of three independent experiments performed on cells of three different donors (n = 9).</p

BrdU incorporation analysis.

<p>Quantification of proliferation rate in hTM after 48 hours normalized to starting activity in controls. (<b>A</b>) Effects of ω-6 (16 µM) and ω-3 (50 µM) fatty acids compared to controls (Co). (<b>B</b>) Effects of H₂O₂ in controls, ω-6 and ω-3 fatty acids pre-treated hTM. (<b>C</b>) %-reduction of BrdU-incorporation after H₂O₂ exposition. Values represent m.a. ± sd of three independent experiments performed in triplicates of 5 different donors (n = 45); asterisks: p-values of statistical significances (*p≤0.05; **p≤0.01; ***p≤0.001).</p

Antibodies used for Western blots (WB) and Immunofluorescence (IF) labeling.

<p>Antibodies used for Western blots (WB) and Immunofluorescence (IF) labeling.</p

Anterior segment OCT scan of lamellar bed (white gas reflections) created by ultraviolet femtosecond laser.

<p>Anterior segment OCT scan of lamellar bed (white gas reflections) created by ultraviolet femtosecond laser.</p