Fig. 1

Production of PAF linked to LPCAT2 activation is enhanced by B. abortus infection. (a) RAW264.7 cells pretreated with AG490 (75 μM) or CV3988 (1 μM), a PAFR antagonist, for 1 h were infected with live B. abortus for 5 min. The cells were infected with heat-killed B. abortus and live E. coli O157:H7 for 5 min. The levels of PAF secretion in the culture supernatants were quantified by PAF ELISA. Data represent the mean ± SD of triplicate trials from three independent experiments. Statistically significant differences from the untreated samples are indicated by asterisks (*, P < 0.05; ***, P < 0.001). (b) Cells pretreated with PAF (200 nM), a PAFR agonist, for 5 min and AG490 (75 μM), a JAK2 inhibitor, for 1 h were infected with B. abortus for 5 min. (c) Cells were also were infected with heat-killed B. abortus or live E. coli O157:H7 for 5 min. The cells were processed for immunoprecipitation with a LPCAT2 antibody and then were probed with a phosphoserine antibody. The membrane was then stripped and re-probed with LPCAT2 antibody. The images shown are representative of three independent experiments