Figure 2
Dose and time course of sPLA 2 -IIA release into BMVEC culture medium. BMVECs were treated with 0, 1, 5, or 10 μg/ml of LPS as described in the Methods, and the cell culture medium was collected to determine the sPLA2-IIA protein and mRNA expression by Western blotting and real-time PCR. The same blotted membrane was stripped and re-probed for BSA, and the relative sPLA2-IIA band intensity was calculated. (A) Representative bands showing the dose response curves of sPLA2-IIA protein (upper panel) and BSA (middle panel) from Western blotting. Quantitative data of sPLA2-IIA release after LPS treatment for 24 h are shown in the lower panel. The data are presented as the means ± SE of the four separate experiments. *P < 0.05 versus the normal group (LPS 0 μg/ml) and LPS treatment with 1 μg/ml. (B) Representative bands showing the Western blotting results for sPLA2-IIA protein (upper panel) and BSA (middle panel). Quantitative data of sPLA2-IIA release after LPS treatment are shown in the lower panel. The data are presented as the means ± SE of the four separate experiments. *P < 0.05 versus the normal group (time 0 h) and LPS treatment for the 8-h group, ^ P < 0.05 versus LPS treatment for the 16-h group. (C) A line chart shows representative results of sPLA2 enzyme activity in the culture medium after 5 μg/ml LPS treatment. The data shown are the averages of four separate experiments from four individual platings (means ± SE). *P < 0.05 versus the normal group (time 0 h) and LPS treatment for the 8-h group, ^ P < 0.05 versus LPS treatment for the 16-h group. (D) Quantitative data of sPLA2-IIA mRNA expression after LPS treatment with 5 μg/ml for different times. The data are presented as the means ± the SE of the four separate experiments. *P < 0.05 versus the normal group (time 0 h), ^ P < 0.05 versus LPS treatment for the 8-h group. (E) Quantitative data of sPLA2-IIA release into the MVECs medium after 5 μg/ml LPS treatment by ELISA. The data are presented as the means ± the SE of the four separate experiments. *P < 0.05 versus the normal group (time 0 h), ^P < 0.05 versus the LPS 8-h group.