Fig. 3

Activation of PAFR-associated JAK2 signaling involved in B. abortus uptake. (a) RAW 264.7 cells were pretreated for 5 min with a serial concentration (0–200 nM) of PAF. Of the PAF-treated cells, cells in 0 and 200 nM PAF were infected with B. abortus for 5 min. The cells were lysed, and the activation of JAK2 and p38α were monitored by immunoblot analysis. Images shown are representatives of three independent experiments. (b) Cells were pretreated with CV3988 (1 μM) for 1 h, followed by infection with B. abortus for the indicated times (5, 15, or 30 min), and then monitored by immunoblot analysis as described in (a). (c) Cells pretreated with or without AG490 (75 μM) for 1 h were stimulated with PAF (200 nM), and then the infection assay was conducted using the same procedure used to evaluate bacterial uptake into macrophages. Statistically significant differences from the untreated samples are indicated by asterisks (**, P < 0.01; ***, P < 0.001)