Figure 4.

Identification of transcription factors interacting with Dnmt1. (A) List of transcription factors interacting with Dnmt1 according to the results obtained from the hybridation of 5 µg of Dnmt1^R on transcription factors arrays (Ozyme, Saint-Quentin-en-Yvelines, France). (B) Validation of the Dnmt1/transcription factors interactions in U251 glioma cells using the P-LISA–Olink/Duolink method. Nucleus/DNA was stained with DAPI (blue), and interactions were visualized by red fluorescence according to the P-LISA–Olink/Duolink method according to a previous report.¹³ (C) Validation of the Dnmt1/transcription factors interactions in U251 glioma cells using the immunoprecipitation method. Immunoprecipitations were realized using the Catch and Release v2.0 Reversible Immunoprecipitation System (Millipore, Molsheim, France). Nuclear extract is obtained using the EpiQuik Nuclear Extraction Kit I (Euromedex, Souffelweyersheim, France). (D) Model of DNA methylation inheritance machinery. Two sides compose the DNA methylation inheritance machinery: one untargeted, represented by the Dnmt1-PCNA-UHRF1 complex, and a second targeted, represented by the Dnmt1/transcription factor interactions. These 2 sides can work in solo or in duet to maintain the DNA methylation level of genes or DNA repeat elements.