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. 2018 Jun 19;32(12):6883–6897. doi: 10.1096/fj.201800413RR

Figure 4.

Figure 4

REDD1 expression is enhanced by consumption of a Western diet and C6-Cer exposure of cells in culture. A) C57BL/6 mice were fed either a Western diet (WD) or control diet (CD) for 14 wk before retinal extraction. REDD1 protein expression was assessed relative to tubulin by Western blot analysis (n = 7). Protein loading was assessed by protein staining (Protein S.). B) R28 cells and MEFs were maintained in culture medium and treated with 1.2 μl/ml DMSO vehicle (Veh) or 60 μM C6-Cer (Cer6). Bands are from the same blot, but are not contiguous. C–F) Quantification of REDD1 protein expression in R28 (C) and MEFs (D) at either 0 or 8 h after addition of Cer6 to the culture medium. REDD1 mRNA expression in R28 cells (E) or in MEFs (F) was assessed by RT-PCR 12 h after addition of either Cer6 or a vehicle to culture medium. Representative blots are shown. Protein molecular mass (kDa) is indicated at the right of the blots. Results are representative of 2 independent experiments; within each experiment, 3 independent samples were analyzed and are means ± sem (n = 3). Different letters (a–c) above scatter plots (A, C–F) denote significantly different results. P < 0.05.