Figure 5.

G. lucidum ethanol extract ameliorated cytotoxicity and apoptosis H₂O₂ induced in HaCaT cells: (a) HaCaT cells were cultured for 6 h in the presence of the indicated concentrations of H₂O₂ (25–800 µM) before MTT assay. The results are expressed as means ± SD of independent experiments performed in triplicate and are reported as percentage vs. the untreated control (ANOVA, * p < 0.05,*** p < 0.001 and **** p < 0.0001 vs. control). (b) HaCaT cells were cultured for 18 h in the presence of the indicated concentrations (0, 5 and 10 µg mL⁻¹) of the G. lucidum ethanol extract before treatment with H₂O₂ for 6 h. The results are expressed as means ± SD of independent experiments performed in triplicate and are reported as percentage vs. the untreated control (ANOVA, * p < 0.05 vs. control). (c) Flow cytometric analysis of annexin V and propidium iodide (PI) double staining in the G. lucidum ethanol extract-treated HaCaT cells after 18 h and H₂O₂ for 6 h: histograms indicate the total percentage of early (annexin V-positive cells/PI-negative cells) and late apoptotic events (annexin V/PI-double positive cells) as well as necrotic cells (annexin V-negative cells/PI-positive cells). The results are representative of four independent experiments performed in duplicate and are expressed as mean ± SD (ANOVA, * p < 0.05).