An Extract from the Plant Deschampsia antarctica Protects Fibroblasts from Senescence Induced by Hydrogen Peroxide

Figure 4

Effect of H₂O₂ treatment in the SA-β-Gal level and protein content of Sirt1 and LmnA/C of HFF cells. (a) The SA-β-Gal level measured as a number of blue cells in control (C13, H₂O₂-unexposed cells) and cells exposed to 100, 150, and 200 μM H₂O₂ during 1 and 2 h and after 24 h of H₂O₂ recovery. Capital letters indicate statistical comparative analysis among data from 1 h H₂O₂ treatment and lowercase letters among 2 h H₂O₂ treatment (b) The β-Gal level measured as nmol of fluorescein per number of cells in controls (H₂O₂-unexposed cells, C13: 13 PD; C26: 26 PD; and C45: 45 PD) and cells exposed to 100, 150, and 200 μM H₂O₂ during 2 h and after 24 h of recovery. (c) Cells stained with the chromogenic substrate 5-bromo-4-chloro-3-indolyl-beta-d-galactopyranoside (X-gal) prior to 100, 150, and 200 μM H₂O₂ exposition and after 24 h of H₂O₂ recovery. (d, e) Densitogram (upper panel) and Western blot for Sirt1 and LmnA/C (lower panel) in controls (H₂O₂-unexposed cells: C13, C45) and H₂O₂-exposed cells during 2 h and after 24 h H₂O₂ recovery. Panel LmnA/C; capital letters indicate statistical comparative analysis among LmnA data and lowercase letters among LmnC. Act-C is used as loading control. Each bar represents the mean ± S.E. of three replicates from three independent experiments, and samples that do not have a common letter are significantly different by Duncan’s test at .

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