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. 2019 Sep 30;129(11):4745–4757. doi: 10.1172/JCI128293

Figure 7. JMJD3 regulates PDLIM4 by facilitating the interaction between KLF2 and WDR5.

Figure 7

(A) 293T cells were cotransfected with HA-Klf2 and FLAG-tagged Ash2L, RbBP5, Wdr5, or Dpy30. WCLs were immunoprecipitated with anti-FLAG beads and immunoblotted with anti-HA Abs. (B) WCLs from WT and Jmjd3-cKO CD4 SP T cells were immunoprecipitated with anti-KLF2 Abs and immunoblotted with anti-JMJD3, WDR5, and KLF2 Abs, respectively. (C) Schematic presentation of cloned HA-tagged N- and C-terminal regions of KLF2. (D) 293T cells were cotransfected with HA–N-terminal, HA–C-terminal, or full-length Klf2 along with FLAG-Wdr5. WCLs were immunoprecipitated with anti-FLAG beads and immunoblotted with anti-HA Abs. (E) 293T cells were cotransfected with HA–N-terminal, HA–C-terminal, or full-length Klf2 along with FLAG-Jmjd3. WCLs were immunoprecipitated with anti-FLAG beads and immunoblotted with anti-HA Abs. (F) 293T cells were cotransfected with HA–N-terminal, HA–C-terminal, or the middle region of Jmjd3 along with HA-Klf2. WCLs were immunoprecipitated with anti-Klf2 Abs and immunoblotted with anti-HA Abs. (G) 293T cells were cotransfected with HA–N-terminal, HA–C-terminal, or the middle region of JMJD3 along with FLAG-Ash2L. WCLs were immunoprecipitated with anti-FLAG Abs and immunoblotted with anti-HA Abs. (H) Schematic diagram of proposed mechanism of how JMJD3 and KLF2 regulate Pdlim4 expression. JMJD3 interacts with the MLL4 protein ASH2L and forms a stable complex with KLF2, which is capable of binding to the Pdlim4 promoter. This permits JMJD3 to alter the H3K27 methylation state of Pdlim4 and permits ASH2L to alter H3K4 methylation to control Pdlim4 expression.