Fig. 2

CsLysoPLA facilitates IL-25 expression in RAW264.7 cells via PKA-dependent B-Raf-ERK1/2 pathway. a RAW264.7 cells were stimulated with CsLysoPLA (5, 10 μg/ml) for 24 h. Relative expressions of PKA, B-Raf, and ERK1/2 genes were examined by Quantitative real-time PCR. b RAW264.7 cells were stimulated with CsLysoPLA (10 μg/ml) in the absence or presence H-89 (20 μM) for 15, 30, 60 and 90 min. The protein levels of phospho-B-Raf, total ERK1/2, phospho-ERK1/2, total AKT, phospho-AKT were detected by western blotting with their respective antibodies. GADPH was used as a loading control. c Quantification of western blot data in (b). “−/−” PBS control, “+/−” cells treated with CsLysoPLA and “+/+” cells treated with CsLysoPLA and H-89. d RAW264.7 cells were stimulated with CsLysoPLA (10 μg/ml) with or without SC79 (4 μg/ml) for 15 and 30 min. The protein levels of total ERK1/2, phospho-ERK1/2, total AKT, phospho-AKT were detected by western blotting with their respective antibodies. PBS was used as negative control. e Quantification of western blot data in (d). “−/−” PBS control, “+/−” cells treated with CsLysoPLA and “+/+” cells treated with CsLysoPLA and SC79. f RAW264.7 cells were stimulated by CsLysoPLA (10 μg/ml) in the presence of H-89 (20 μM) or SC79 (4 μg/ml) for 12 h. The level of IL-25 mRNA was analyzed by quantitative real-time PCR. Data are shown as mean ± SEM. *P < 0.05