Fig. 2.
Effects of mushroom extract number 28 on UVB-induced senescence in HaCaT cells. a Identification of mushroom extracts that augment SIRT1 transcription in HaCaT cells. HaCaT cells transduced with vector expressing EGFP under the control of the SIRT1 promoter (2.0 × 104 cells) were seeded on to 96-well μClear Fluorescence Black plate and cultured for 24 h. Mushroom extracts (10 μg/mL) were added and changes in EGFP fluorescence were monitored using an IN Cell Analyzer 1000. b Effects of mushroom extracts on the expression of endogenous SIRT1 in HaCaT cells. HaCaT cells were treated with 10 μg/mL of mushroom extracts. Next day, RNA was prepared and quantitative RT-PCR was conducted. The results are expressed as mean ± SD. Statistical significance was defined as *p < 0.05, ***p < 0.001. c Effects of mushroom extract number 28 on UVB-induced senescence in HaCaT cells. Cells were firstly treated with resveratrol and mushroom extract number 28 (10 μg/mL) every 3 days, irradiated with 10 mJ/cm2 UVB in the presence of extract, and cultured for 3 days. Senescence-related biomarkers, including SA-β-Gal activity, γH2AX, p16, p21, and phospho-p38 MAP kinase, were detected using an IN Cell Analyzer 1000. Resveratrol (5 μM) was used as a positive control. The relative number of cells with high activity or expression of senescence biomarkers is shown in black in the pie chart. Multiple comparisons against non-treated HaCaT (#) or against UVB-treated HaCaT ($) were calculated by one-way ANOVA with Turkey’s post hoc test. Significant differences are denoted by ###, $$$ p < 0.001