Figure 5

Immunofluorescence staining for TNF-R1; a) in the fresh, organ culture after 24 h incubation, organ culture after 48 h incubation, 24 h culture + treatment with IMD-0354, 48 h culture + treatment with IMD-0354, 24 h culture + treatment with SB386023-b, 48 h culture + treatment with SB386023-b, 24 h culture+ treatment with U0126 and 48. There was a marked increase in TNF-R1 protein level in the smooth muscle cell layer after 24 and 48 h culture as compared to fresh vessels. TNF-R1 was significantly decreased with all three inhibitors at 48 h of organ culture and reduced at 24 h. b) Bar graph demonstrating the fluorescence intensity for TNF-R1. The results are expressed as mean values ± S.E.M. *P < 0.05 and **P < 0.01, significant difference between fresh and culture after 24 and 48 h. ^§P < 0.05 and ^§§P < 0.01, significant difference between 48 h organ culture and treatment with 48 h culture + (IMD-0354, SB386023-b and U0126). ^#P < 0.05 and ^##P < 0.01, significant difference between 24 h organ culture and treatment with 24 h culture + (IMD-0354, SB386023-b and U0126). n.s. = not significant.