Fig. 5

WDR13 interacts with c-Jun and binds at AP1 sites of c-Jun promoter. a Immunoprecipitation, using anti WDR13 antibody from mouse embryonic fibroblasts shows interaction of WDR13 with c-Jun. Wdr13 knockout MEFs were used as control. Arrows show specific band. * Denotes non-specific band in input and IgG band in IP samples.b Co-immunoprecipitation of Myc-c-Jun and FLAG-WDR13 using anti FLAG agarose beads in HEK293 cells show interaction of WDR13 with c-Jun. Interaction of WDR13 with c-Jun is enhanced as phosphorylation status of c-Jun increases. c Domain organization of WDR13 and schematic of various deletion constructs. d Domain organization of c-Jun and schematic of various deletion constructs. e Co-immunoprecipitation of Myc-c-Jun and various deleted FLAG-tagged WDR13 domains using anti FLAG agarose beads in HEK293 cells shows that region of 293–393 amino acids is essential for interaction. 5% input shows protein expression in cell lysate with Myc specific antibody in the lower panels. f Co-immunoprecipitation of FLAG-Wdr13 and various deleted Myc tagged c-Jun domains using anti FLAG agarose beads in HEK293 cells shows interaction of WDR13 with DNAdimerization and DNA-binding domain of c-Jun. 5% input shows protein expression in cell lysate with WDR13-specific antibody in the lower panels. g Schematic overview of c-jun promoter. Primer pairs used for ChIP from AP1 site were underlined. h WDR13 binding to AP1 site of c-Jun promoter in MEFs. Wdr13 knockout MEFs was used as ChIP control. White bar, Wdr13 ^-/0 MEFs 2 h after UV treatment; black bar, Wdr13 ^+/0 MEFs; Light grey bar, Wdr13 ^+/0 MEFs 2 h after UV treatment. ChIP experiment was performed from independently derived two MEFs line for each genotype. Student’s t-test was used for statistical analysis and error bar shows s.e.m