Fig. 4

ZNF300ZFR mediates DNA methylation. a A co-immunoprecipitation assay was performed in HEK293T cells co-transfected with Flag-ZNF300 and HA-KAP1 to examine the interaction between ZNF300 and KAP1. b Schematic illustrations show the position of CpG sites (circles). The arrow indicates the transcription start position and numbers indicate the position of CpG sites relative to transcription start site (upper). Bisulfite sequencing analysis the methylated CpGs in the ZNF300 promoter (ZNF300pro) with ZNF300 ZFR on the downstream as described in Fig. 1d. Black circles indicate methylated CpGs and open circles indicate non-methylated CpGs (lower). c K562 cells were treated with vehicle or 5-AzadC. The ZNF300 protein was measured by Western blot (left panel) and the ZNF300 mRNA expression was measured by quantitative RT-PCR (right panel). d K562 cells treated with PMA or vehicle as indicated for 2 days. The CD61 expression of the resultant cells was measured by staining cells with anti-CD61-PE antibody and analyzed by flow cytometry. e K562 cells were treated with vehicle, PMA, 5-AzadC, or PMA/5-AzadC and the CD61 expression were measured by staining cells with an anti-CD61-PE antibody and analyzed by flow cytometry. f ZNF300 knockdown cells (shZNF300 #2 or #4) were treated with PMA/5-AzadC as indicated. The CD61 expression of the resultant cells were measured by staining cells with an anti-CD61-PE antibody and analyzed by flow cytometry