Fig. 1.

LPS triggers Alcat1 degradation. (A–C) MLE cells were treated with LPS at varying concentrations (B, lower panels) or times (B, upper panel; fixed LPS concentration of 4 µg/ml). The oxygen consumption rate of cells was analyzed with a seahorse XF analyzer (A), or cell lysates were immunoblotted for Alcat1 and β-actin (B). (C) C57BL6 mice were treated with LPS (5 mg/kg) intratracheally overnight, and whole-lung tissue lysates were analyzed by immunoblotting for Alcat1 and β-actin. (D) Total cellular RNA was isolated from untreated or LPS-treated MLE cells in B (4 µg/ml for 16 h), and quantitative PCR analysis was conducted to determine steady-state mRNA levels using Alcat1-specific primers. Cox2 was used as a positive control. *P<0.05, Cox2 mRNA changes in untreated versus LPS-treated cells (Student's t-test). Data in each panel represents n=3 separate experiments. Means±s.e.m. are shown.