Figure 7
Indirubin derivatives E804 and E231 inhibit H9N2-induced p38 and JNK signaling pathways in human pulmonary endothelial cells.
(A,B) HPMECs were infected with H9N2 at 2 MOI followed by treatment with indirubin derivatives E804 or E231 (1 μM). Total cell lysates were harvested at 24 h.p.i. (for p-p38) or 6 h.p.i. (for p-JNK). Phosphorylation and total p38 and JNK were detected by Western blot analysis with specific antibodies. Representative image from three independent experiments (Upper panel). Quantitative analysis of Western blotting. Values are presented as fold-change of phosphorylated p38 or JNK normalized to corresponding total form and then compared with mock-infected cells (Lower panel). (C,D) E804 directly inhibits p38 and JNK kinases activity in vitro. HPMECs were infected with H9N2 at 2 MOI. Cell lysates were harvested at 24 h.p.i. (for p-p38) or 6 h.p.i. (for p-JNK). In vitro kinase assay of p38 and JNK was then performed as described in materials and methods. Representative image from three independent experiments (Upper panel). Quantitative analysis of Western blotting. Values are presented as fold-change of phosphorylated ATF2 or c-Jun compared with mock-infected cells (Lower panel). The values are presented as mean ± S.D. of three independent experiments. **p < 0.01, ***p < 0.001 vs mock-infected cells, #p < 0.05 vs H9N2-infected cells.
