Figure 5.

BCA2 facilitates MyD88 interaction with TLR4 and inhibits MyD88 interaction with OTUD4. (A) HEK293T cells were transfected with different GST-fused MyD88 fragments and Flag-OTUD4. After 48 hours of transfection, cells were harvested for GST pull-down. (B) In HCC1806 cells with BCA2 knockout, MyD88 was immunoprecipitated with endogenous OTUD4. (C) In HCC1806 cells overexpressing BCA2, MyD88 was immunoprecipitated with endogenous OTUD4. (D) Endogenous ubiquitination experiments were performed in HCC1806 cells with BCA2 knockout after stimulation with LPS (500 ng/mL) using a ubiquitin Ab. (E) HCC1806 cells with BCA2 knockout were stimulated with LPS (500 ng/ml) for 0-30 minutes before immunoprecipitation and immunoblot analysis.