Fig 1: Cortisol triggered GR-dependent downregulation of Rer1 (A) A thousand base pair upstream of the first codon of the RER1 was described and the putative GRE binding sequence was emphasized with yellow labeling. (B) SH-SY5Y cells were incubated with cortisol for 6 h (1 µM). DNA was immunoprecipitated with RNA Pol, IgG, and GR antibody. The immunoprecipitation and input samples were amplified with primers of GAPDH and RER1. n = 5. (C – F) SH-SY5Y cells were transfected with NT siRNA or GR siRNA for 24 h, and treated with cortisol during 12 h (1 µM). (C) The expressions of Rer1 and ß-actin were detected by western blotting. ß-actin was used as a loading control. n = 5. (D) TGN38 was co-immunoprecipitated with Rer1. Rer1 levels in immunoprecipitated samples were quantified. n = 5. (E) The cells were immunostained with TGN38 (green), DAPI (blue), and Rer1 (red). Pearson’s correlation coefficient between TGN38 and Rer1 was quantified. Scale bars, 20 µm (magnification, × 1,000). n = 5. (F) The cells were immunostained with Rab7 (green), DAPI (blue), and PSEN1 (red). Pearson’s correlation coefficient between Rab7 and PSEN1 was quantified. Scale bars, 20 µm (magnification, × 1,000). n = 5. All blots and immunofluorescence images are representative. Quantitative data are presented as a mean ± S.E.M. The representative images were acquired by SRRF imaging system. ** indicates p<0.01 versus control. ## indicates p<0.01 versus cortisol, respectively. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)