Skip to main content
. 2017 Oct 3;3:17035. doi: 10.1038/celldisc.2017.35

Figure 7.

Figure 7

YY2 somatic mutations alter K247 methylation, DNA-binding activity and its regulated gene transcription. (a) HeLa cells transfected with vector expressing Flag-tagged YY2 (wt), YY2 (K244Q) or YY2 (S246F) were subjected to IB with antibodies as indicated. Intensity of YY2K247me1 was quantified using Image J and is shown as indicated. (b) DNA EMSA assay was performed as described in Figure 4a with whole-cell lysates prepared from HeLa cells transfected with vector expressing Flag-tagged YY2 (wt), YY2 (K244Q) or YY2 (S246F). Intensity of shifted band was quantified by using Image J and shown as indicated. (c) HeLa cells were transfected with pGL2-YY1-luc vector and vectors expressing HA-tagged YY2 (wt), YY2 (K244Q) or YY2 (S246F), followed by ChIP and qPCR as described in Figure 4h. ChIP signals were presented as fold induction over wt after being normalized to input (±s.e.m., ***P<0.001). (d) HeLa cells were transfected with vectors expressing HA-tagged YY2 (wt), YY2 (K244Q) or YY2 (S246F), followed by ChIP as described in Figure 5d. ChIP signals were presented as fold induction over wt after being normalized to input (±s.e.m., ***P<0.001). (e, f) HeLa cells were transfected with control vector or vectors expressing YY2 (wt), YY2 (K244Q) or YY2 (S246F), followed by RT-qPCR analysis to examine mRNA levels of selected genes as indicated. Data shown were the relative fold change compared with control samples after normalization to actin (±s.e.m., *P<0.05, **P<0.01, ***P<0.001; NS, nonsignificant).