Figure 6.

Geometric constraints control the subcellular distribution of DNMT1 in a PI3K/AKT‐dependent manner. A) Representative immunofluorescence staining and quantification of the colocalization of DNMT1 and AKT1. Images were acquired and processed using stimulated emission depletion super‐resolution microscopy. Correlation is numerically shown using the Pearson's coefficient. B,C) Micropatterned SMCs were treated with a pan‐AKT inhibitor (GSK690693, 10 µmol L⁻¹), PI3K inhibitor (LY294002, 10 µmol L⁻¹), or control reagent (DMSO) for 24 h; representative immunofluorescence images and quantifications of the nuclear/cytoplasmic ratio of DNMT1 are shown. Fluorescence images were acquired using an epifluorescence microscope. In (A)–(C), each dot represents a single cell. D,E) Micropatterned cells were treated with an AKT inhibitor (GSK690693), PI3K inhibitor (LY294002), or a control reagent (DMSO), and representative results of the agarose gel electrophoresis of the products of MSP for the D‐loop of bisulfite‐modified DNA isolated from the cells are shown. Semi‐quantification of the MSP results is presented in the lower panels. M: methylated, U: unmethylated. Data were obtained from four biological repeats. Significance was assessed using A,D,E) a Student's t‐test and B,C) two‐way ANOVA with Tukey's post hoc analysis. The error bars show ± SD. The exact P values between the indicated groups are presented.