Figure 5. Jmjd3 Interacts with PHF20 and Causes its Degradation.
(A) 293T cells were transfected with Flag-PHF20 and HA-Jmjd3. Cell extracts were immunoprecipitated with anti-Flag beads, followed by immunoblot (IB) with an anti-HA antibody. (B) Detection of interaction between Jmjd3 and PHF20 in WT and PHF20 knockout MEFs by immunoprecipitation (IP) with a PHF20 antibody, followed by IB with an anti-Jmjd3 antibody. (C and D) 293T cells were transfected with different HA-Jmjd3 domain constructs (C) and different Flag-PHF20 constructs (D). Cell extracts were immunoprecipitated with anti-Flag beads, followed by IB with an anti-HA antibody. (E) 293T cells were transfected with increasing amounts of HA-Jmjd3 cDNA, followed by IB with anti-PHF20 and anti-HA antibodies. (F) Jmjd3 WT and KO MEF cells were infected with or without Flag-tagged Jmjd3 retrovirus for24 h, and cell extracts were analyzed by IB with anti-PHF20 and anti-Jmjd3 antibodies. See also Figure S5.