FIG. 4.

Time course effects of AMS on keratin expression in HaCaT cells. (A) Western blot analysis for the evaluation of the keratin total content in HaCaT cells exposed for 24 and 48 h to 0.46 mg/mL of AMS (Annurca apple polyphenolic extract microencapsulated with maltodextrins, AppleMetS), in the presence (left blot) or absence (right blot) of serum. Equal amounts of proteins (70 μg) were separated on a 10% SDS–polyacrylamide gel and subjected to Western blot analysis using monoclonal anti-pan-cytokeratin antibodies that recognize the following human cytokeratins according to their molecular weight: K1 (68 kDa), K4 (59 kDa), K5 (58 kDa), K6/K10 (56 kDa), K13 (54 kDa), K8 (52 kDa), K18 (45 kDa), and K19 (40 kDa). Shown are blots representative of three independent experiments. (B) The cytokeratin bands within each lane were quantified by densitometric analysis and plotted in line graph as percentage of control. The anti-GAPDH antibody was used to standardize the amounts of proteins in each lane. Shown are the averages ± SEM values of three independent experiments. ***P < .001 versus control (untreated cells).