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. 2021 Oct 8;26(19):6074. doi: 10.3390/molecules26196074

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© 2021 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Effects of AFW4 on cell proliferation and skin hydration are mediated by activating p44/42 MAPK. (A–C) HaCaT cells were co-transfected with the AP-1-, NF-κB-, or CRE-promoter-luciferase reporters and β-galactosidase reporter vector using polyethylenimine. After 24 h, the transfected cells were incubated with AFW4, phorbol 12-myristate 13-acetate (PMA), tumor necrosis factor (TNF)-α or forskolin (Fk). Twenty-four hours after the incubation, the cells were harvested and subjected to luciferase reporter assay. (D) Cells were incubated with AFW4 or PMA for 1 h. The cells were harvested immediately after the incubation, and the protein levels of MAPKs and their phosphorylated forms were detected by ELISA. The results were confirmed using four independent experiments. Each experiment was conducted in duplicate. All data are presented as the mean ± SEM of four independent experiments. Statistical significance of differences among the groups was assessed by ANOVA, followed by Tukey’s multiple-comparison test, using the GraphPad Prism 5 software. * p < 0.05 vs. control group.

Effects of AFW4 on cell proliferation and skin hydration are mediated by activating p44/42 MAPK. (A–C) HaCaT cells were co-transfected with the AP-1-, NF-κB-, or CRE-promoter-luciferase reporters and β-galactosidase reporter vector using polyethylenimine. After 24 h, the transfected cells were incubated with AFW4, phorbol 12-myristate 13-acetate (PMA), tumor necrosis factor (TNF)-α or forskolin (Fk). Twenty-four hours after the incubation, the cells were harvested and subjected to luciferase reporter assay. (D) Cells were incubated with AFW4 or PMA for 1 h. The cells were harvested immediately after the incubation, and the protein levels of MAPKs and their phosphorylated forms were detected by ELISA. The results were confirmed using four independent experiments. Each experiment was conducted in duplicate. All data are presented as the mean ± SEM of four independent experiments. Statistical significance of differences among the groups was assessed by ANOVA, followed by Tukey’s multiple-comparison test, using the GraphPad Prism 5 software. * p < 0.05 vs. control group.