Figure 5.

Role of Dux and Duxbl1 on Zscan4 promoter regulation. (A) Schematic representation of Dux and Duxbl1 protein domains. (B) Dux and Duxbl1 expressions were significantly induced by RA. The normalization was performed using Gapdh as housekeeping gene. (top). ES^Zscan4_Em cells grown for 5 days with or without RA and sorted by flow cytometry. Dux and Duxbl1 expressions were robustly enriched in Zscan4⁺ cells (bottom). The normalization was performed using Gapdh as housekeeping gene. (C) HEK293T cells with Luciferase-reporter gene under Zscan4 promoter regulation (scheme) were transiently co-transfected with: control Vector, or Dux-HA, Duxbl1-Flag either alone or with Dux-HA. Data shown represent relative luciferase activity normalized against the β-galactosidase activity, in the top panel, while in the bottom panel was reported the western blot assay corresponding to luciferase assay. Dux-HA and Duxbl1-Flag were detected with anti-HA, anti-Flag antibodies while anti-Gapdh was used for proteins normalization. *p < 0.05, ***p < 0.01, ***p < 0.001 in a Student's t-test.