Fig 1: PI acyl chain profile regulates TIRAP expression.(A) Control and LPS-primed WT cells grown on coverslips were labeled with anti-TIRAP Ab, and nuclei were stained with DAPI. The plasma membrane was stained by adding Alexa Fluor 647–conjugated phalloidin for the last 15 min. (B) WT, Abcb1–/–, and LA-supplemented cells were stimulated with LPS for different times. Cell lysates were collected and immunoblotted for TIRAP and GAPDH. (C) Quantitation of the WB shown in (B) by ImageJ. (D and E) WT (D) and LA-grown (E) cells were pretreated or not with the proteasomal inhibitor MG132 (10 μM) for 30 min before stimulating the cells with LPS for different time points. Cell lysates were collected and immunoblotted for TIRAP and GAPDH. (F and G) WT, Abcb1b–/–, and LA-supplemented cells were exposed to IL-1 (2 ng/ml) overnight. RNA was extracted and converted into cDNA. Gene expression of Nlrp3 and TNF-α was determined by real-time PCR. The data shown are representative of at least three independent experiments with three to five replicates each. Asterisk (*) on immunoblots denotes a nonspecific band. Scale bars, 5 μm.