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MEL-18 controls ESR1 transcription by suppressing the new SUMOylation of the ESR1 transcription factors p53 and you may SP1
(A) Cell lysates treated with 20 mM N-ethylmaleimide (NEM) were subjected to immunoblotting. The amount of SUMOylated protein was quantified by measuring the ratio of SUMOylated protein/total protein. (B) Venn diagram showing the relationship between the microarray results for MCF-7 cells expressing MEL-18 shRNA (shMEL) and those for MCF-7 cells treated with RITA (GSE13291) ( 36 ). (C) MCF-7 cells expressing MEL-18 siRNA (siMEL) were cotransfected with WT or SUMOylation-deficient mutant constructs of p53 or SP1 and with ESR1 pro-Luciferase and were subjected to a luciferase reporter assay. The data are presented as the mean ± site web de rencontres pour célibataires de la musique SD (n = 3). *P < 0.05 vs. siCon/Con; † P < 0.05 siMEL/Con (2-tailed Student's t test). (D) ChIP-qPCR analysis showing the amount of ESR1 transcription factor that was recruited to the ESR1 promoter in the indicated cells. The data are presented as the mean ± SD (n = 3). *P < 0.05 vs. shCon (2-tailed Student's t test). (E) The effect of ginkgolic acid on the expression of ER-? in the MEL-18–silenced cells. Cells were treated with 100 mM ginkgolic acid for 24 hours and subjected to immunoblotting. Parallel samples examined on separate gels are shown. The data were quantified by measuring the immunoblot band densities from three independent experiments (mean ± SD). *P < 0.05 vs. shCon; † P < 0.05 vs. shMEL (2-tailed Student's t test). All data shown are representative of three independent experiments.
In the MEL-18–silenced MCF-7 structure, the amount of the 39-kDa SUMO-1–conjugating particular the brand new SUMO E2 enzyme UBC9 was enriched, while the level of the fresh new 18-kDa free-form away from UBC9 is less (Extra Shape 13A)
MEL-18 advances deSUMOylation from the inhibiting new ubiquitin-proteasome destruction off sentrin-certain protease step one. To further pick new apparatus for which MEL-18 controls SUMOylation, the outcome of MEL-18 into term regarding SUMO-related circumstances try checked.
