Figure 1.

Identification of IRAK1-binding partners by yeast two-hybrid screening. A, AH109 cells were transfected with each of the pGADT7-based putative positive clones, which are mentioned in Table 1 and also with pGBKT7-based IRAK1 or p53. Upper and lower plates are SD/−leucine/−tryptophan and SD/−leucine/−tryptophan/−histidine/+3-AT, respectively. B, a truncated form of OPTN (OPTN425–577) was cloned and expressed in yeast cells. C, HEK293T cells were transfected with FLAG-tagged IRAK1 or IRAK1-KD along with Myc-tagged OPTN, OPTN425–577, or TRAF6. After 48 h of transfection, immunoprecipitation was performed using anti-FLAG antibody. Then Western blotting (WB) was performed using anti-Myc or anti-FLAG antibody. The data are representative of three independent experiments with similar results.