Figure 5.

In cellula interaction of CXXC5 and MeCP2. (a) Proximity ligation assay (PLA). To assess the in cellula interaction of CXXC5 and MeCP2, HEK293 cells grown in coverslips were transiently co-transfected the expression vector bearing the 3F-CXXC5 or HA-MeCP2 cDNA. Cells were fixed, permeabilized, blocked, and probed with the HA and/or the Flag antibody. Cells were then subjected to fluorescent probes for circular DNA amplification. DAPI was used for nuclear staining. The scale bar is 25 µm. (b) Chromatin immunoprecipitation assay (ChIP)-WB. Co-transfected cells were also subjected to ChIP using the Flag antibody or the isotype-matched IgG followed by immunoblotting using the HA antibody. The membrane was also re-probed with the Flag antibody. HC and LC indicate the heavy and light chain of IgG. 10% of ChIP was used as input control.