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. 2017 Mar 21;18(5):809–825. doi: 10.15252/embr.201643617

Figure EV3. CUEDC2 regulates GLUT3 via GR .

Figure EV3

  1. Immunoprecipitation (IP) assay was performed with anti‐FLAG antibody in 293T cells cotransfected with HA‐CUEDC2, FLAG‐GR, or FLAG‐EV, followed by blotting with anti‐CUEDC2 or anti‐GR.
  2. Protein levels of GR and GLUT3 were determined by Western blot in HeLa cells and MDA‐MB‐231 cells expressing shCUEDC2 or overexpressing HA‐CUEDC2.
  3. mRNA levels of CUEDC2 and GR were detected by qRT–PCR in PLC cells stably expressing shCUEDC2.
  4. The protein level of GLUT3 in PLC cells stably expressing shGRs was analyzed by Western blot.
  5. mRNA and protein levels of GLUT3 were determined by qRT–PCR and Western blot in PLC cells stably expressing shCUEDC2 with further overexpression of EV or HA‐GR.
  6. Cellular ROS levels were detected by flow cytometry using CellROX DeepRed staining in PLC cells stably expressing CUEDC2 with further knockdown of GR by shRNAs.
Data information: (C and E) Data are presented as mean (± SD); n = 3 in each group. *P < 0.05 as compared to NTC group in (C), and to NTC + EV or to NTC + GR group in (E), respectively. P was calculated by Student's t‐test. NS: Not significant between indicated groups. The representative results of three independent experiments are shown in (F). β‐Actin served as loading control. Source data are available online for this figure.