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. 2014 Dec 11;7(1):42–58. doi: 10.15252/emmm.201404601

Figure 2. REDD1 protein expression in epidermis is tightly regulated and correlates with inhibition of mTOR and autophagy.

Figure 2

B6D2 mice were treated topically as in Fig1. Skin was harvested 4–24 h after single FA application or during chronic treatment (8 h after 1st, 2nd, and 4th applications).
  • A Immunohistochemical staining of mouse skin treated with acetone or FA for 8 h for REDD1 (upper panels) and phosphorylated mTOR-Ser2448 (lower panels). Scale bars are 20 μm.
  • B Western blot analysis of REDD1 protein and phosphorylation of down-stream mTOR target proteins 4E-BP1 and rpS6 in murine epidermis. GAPDH is used as a normalization control.
  • C, D Glucocorticoids induce autophagy in epidermis. Western blot analysis of Beclin-1 and conversion of light chain 3 (LC3) from LC3-I to LC3-II (C). Quantification of LC3-I to LC3-II conversion and Beclin-1 expression (D). The means ± SD were calculated using Western blots from two independent experiments (each lane is whole-cell protein from three pulled individual samples of epidermis).