Fig. 5

Silencing IRAK1 alleviates the production of inflammatory factors caused by BAP31 deficiency. a Scramble and shBAP31 BV2 cells were transfected with IRAK1 siRNA for 60 h, followed by treatment with LPS for 30 min. The cytosolic and nuclear fractions were analyzed by Western blotting with antibodies to c-Jun, p65, histone, and β-actin. b Immunoblots for c-Jun and p65 in cytosolic fractions were quantified and normalized to β-actin protein; immunoblots for c-Jun and p65 in nuclear fractions were quantified and normalized to histone protein. c Scramble and shBAP31 BV2 cells were treated with IRAK1 inhibitor for 48 h, followed by stimulation with LPS for 24 h. The secreted protein levels of the cytokines IL-1β and TNFα in supernatant were analyzed with ELISA kits. d, e Scramble and shBAP31 HEK293T cells were transfected with pcDNA3.1(−) and BAP31-flag plasmids, respectively, for 48 h. Cell lysates were analyzed by Western blotting with antibodies to IRAK1, BAP31, and β-actin. All the data are indicated as Mean ± SEM of three independent experiments. *P < 0.05; **P < 0.01; ***P < 0.001 versus control group