Fig. 3. siRNA-mediated silencing of silent mating type information regulation 2 homolog (SIRT1) blocks Acrp30 stimulation and filaggrin (FLG) regulation. Human normal epidermal keratinocytes were transfected with 20 nM of SIRT1-targeting siRNA or scrambled siRNA and incubated for 48 h. Next, the transfected cells were treated with 10 µg/ml Acrp30 for 24 h. Cell lysates were generated with described lysis buffer and analyzed by immunoblotting. The protein amounts were quantified by comparison to β-actin. Immunoblotting was used to analyze the levels of SIRT1, aryl hydrocarbon receptor nuclear translocator (ARNT), and FLG. All blots shown are representative of 3 independent experiments. ^**p<0.05 compared to the untreated group,. ^***p<0.001 compared to the untreated group, ^****p <0.0005 compared to the untreated group.