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Fig. 4 | BMC Genomics

Fig. 4

From: Co-activator candidate interactions for orphan nuclear receptor NR2E1

Fig. 4

NR2E1 variants R274G and R276G exhibited decreased binding to atrophin-1. GST, or GST-Atrophin-1846–1191 containing the Atro box, were over-expressed in E.coli and incubated with HEK293 cell lysate containing either FLAG-NR2E1LBD, FLAG-NR2E1LBD R274G, or FLAG-NR2E1LBD R276Q. After incubation with glutathione/sepharose beads, purified complexes were resolved by SDS-PAGE followed by western blot using an anti-FLAG antibody. a Note the decreased binding of the FLAG-NR2E1LBD R274G and R276Q variants to GST-Arophin-1846–1191. b Western blot quantification of three independent experiments. The signal intensity detected with anti-FLAG antibody for each pull down was normalized to the input signal. Note that wild-type NR2E1 binds eight times more to atrophin-1 compared to R274G. Error bars, standard error of the mean; GST, glutathione-S-transferase; LBD, ligand binding domain FLAG. n = 3

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