Figure 5
From: Proximity ligation in situ assay for monitoring the global DNA methylation in cells
Impact of conditions increasing the level of DNA methylation on the number of Dnmt1/PCNA interactions. (A) Characterization cell cycle of arrest. U251 cells were treated by serum deprivation (72 h) or by thymidine (2 mM, 24 h), previous to perform cell cycle analysis by using the NucleoCounter NC-3000™ Kit, Chemometec, France). (B) Dnmt1/PCNA P-LISA in U251 cells blocked in G0/G1 or S phases of the cell cycle. Red dots symbolize Dnmt1/PCNA interactions. Nucleus/DNA are stained in blue via the use of DAPI. (C) Analysis of the number of Dnmt1/PCNA interactions (P-LISA) and the 5 mC number (ELISA). The number of Dnmt1/PCNA interactions is calculated from the analysis of 100 nuclei. (D) Impact of the Dnmt3a overexpression in U87 cells on the level of methylation of the capase-8 gene (MeDIP), on DNA (5 mC ELISA) and on the number of Dnmt1/PCNA interactions. The number of Dnmt1/PCNA interactions is calculated from the analysis of 100 nuclei. U87-pORF-Dnmt3a and the U87-pORF (transfection control (-)) were obtained as previously described [16].