Figure 1.

Glyphosate exposure promotes a TET3-mediated global DNA hypomethylation. MCF10A cells were treated according to a timetable shown in (A) and analyzed on day 21 of culture. (Explanations for color-coded days are located in corresponding color rectangles underneath the timeline. UP peptide promotes DNMT1/PCNA/UHRF1 disruption). (B) ELISA was used to measure the level of 5-methylcytosine (5-mC). (C) DMB assay was used to measure maintenance methyltransferase (mMTase). (D) TET assay. (E) In-Cell ELISA was used to quantify TET proteins. (F) MCF10A cells were transfected either with siRNA for TET3 or with control siRNA (siRNA-A) and treated with glyphosate (Glyphosate) or vehicle DMSO (MCF10A) according to a timetable shown in (A). ELISA was used to measure the level of 5mC, and TET3 levels were determined by In-Cell ELISA and normalized to Janus Green staining intensity to account for differences in cell seeding density. For all assays, the bar graph displays the average ± standard deviation values of three independent experiments.