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. 2019 Jul 1;20(3):2294–2302. doi: 10.3892/mmr.2019.10450

Figure 3.

Figure 3.

ATRA decreases Hrd1 expression and increases Nrf2 expression in human fibroblasts exposed to UV radiation. Human fibroblasts were exposed to UVA and UVB, and then treated with ATRA for 24 h. (A) The mRNA level of Hrd1 was assessed using real-time RT-PCR and (B) the protein level of Hrd1 was assessed using western blotting. (C) The mRNA level of Nrf2 was assessed using real-time RT-PCR and (D) the protein level of Nrf2 was assessed using western blotting. (E) Fibroblasts were pretreated with MG132 (10 µM) for 6 h and the endogenous protein-protein interactions between Hrd1 and Nrf2 were assessed by IP with Hrd1 antibody (upper image) or Nrf2 antibody (lower image), followed by IB with Hrd1 antibody and Nrf2 antibody. IgG was used as a negative control for IP. (F) Representative images of Hrd1 interaction with Nrf2. Fibroblasts were stained with Hrd1 (green) and Nrf2 (red). Hrd1 and Nrf2 merged appeared as orange/yellow. Compared to control, **P<0.01; compared to UV; ##P<0.01. ATRA, all-trans retinoic acid; Hrd1, 3-hydroxy-3-methylglutaryl reductase degradation; Nrf2, NF-E2-related factor 2; UV, ultraviolet; IP, immunoprecipitation; IB, immunoblotting.