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MED15 binds and regulates cadmium and zinc responsive genes in mammalian cells.

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posted on 2019-12-09, 18:31 authored by Naomi Shomer, Alexandre Zacharie Kadhim, Jennifer Margaret Grants, Xuanjin Cheng, Deema Alhusari, Forum Bhanshali, Amy Fong-Yuk Poon, Michelle Ying Ya Lee, Anik Muhuri, Jung In Park, James Shih, Dongyeop Lee, Seung-Jae V. Lee, Francis Christopher Lynn, Stefan Taubert

[A] qRT-PCR analysis of MED15 mRNA in control (scr = scrambled siRNA) and MED15 siRNA (siMED15) treated A549 cells exposed to 5μM cadmium for four hours. Error bars: SEM; statistical analysis: * p < 0.05, One-way ANOVA, multiple comparison, Tukey correction, all comparisons to “scr” (n = 3 independent repeats). [B] qRT-PCR analysis of cadmium inducible genes in control (scr = scrambled siRNA) and MED15 siRNA (siMED15) treated A549 cells exposed to 5μM cadmium for four hours. Error bars: SEM; statistical analysis: * p < 0.05, ** p < 0.01, Two-way ANOVA, multiple comparisons, Tukey correction (n = 3 independent repeats). MT1X expression was not significantly affected. For MT2A, all comparisons are vs. “scr”, except where indicated by lines. [C] The graph shows relative MED15 occupancy at the MT1X and MT2A promoters and nearby control regions in A549 cells, as determined by ChIP-qPCR before and after the addition of 5μM cadmium. Error bars: SEM; statistical analysis: ** p < 0.01 Student’s t-test, multiple comparisons, Holm-Sidak correction (n = 3 independent repeats). [D] The graph shows qRT-PCR analysis of MIN6 cells treated without and with 50μM zinc. Error bars: SEM; statistical analysis: ** p < 0.01, *** p < 0.001 Student’s t-test, multiple comparisons, Holm-Sidak correction (n = 4 independent repeats). [E] The graph shows relative Med15 occupancy at the Mt1 promoter and Slc30a8 enhancer and nearby control regions in MIN6 cells as determined by ChIP-qPCR, before and after the addition of 100μM Zn. Error bars: SEM; statistical analysis: * p < 0.05, *** p < 0.01 Student’s t-test, multiple comparisons, Holm-Sidak correction (n = 3 independent repeats).

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