Figure 4: AK023948 is required for the interaction between DHX9 and p85.

(a) AK023948 KO primarily suppresses the p85β level, as detected by western blot. (b) AK023948 interacts with p85, as detected by RIP assay using p85 antibody. (c) AK023948 is required for the interaction between DHX9 and p85, as detected by co-immunoprecipitation (co-IP) using p85 antibody. HMLE cells express little AK023948, whereas MCF-7 cells express a high level of AK023948. (d) Rescue experiments further suggest that AK023948 is critical for the interaction between AK023948 and p85. (e) AK023948 is required for the interaction between DHX9 and p85, as determined by GST pulldown assay. The DHX9 level pulled down by GST-p85 was lower in KO cells than in gRNA control. (f) AK023948 is required for the interaction between DHX9 and p85, as detected by the Duolink in situ Fluorescence Kit (Sigma). HeLa cells were transfected with Myc-p85 plus control siRNA or AK0 siRNA. After 48 h, the cells were fixed for PLA. The red signals were lower in AK0 siRNA than in control siRNA cells. Scale bar, 100 μm. (g) DHX9 siRNAs suppress, but ectopic expression of DHX9 increases both p85 and pAKT. Values in b are s.e.m. (n=3). *P<0.05 by two-tailed Student’s t-test.