Figure 1.

Constitutive expression of LT pathway enzyme mRNA and protein in human bronchial fibroblasts. (A) RT-PCR analyses of human bronchial fibroblasts (F) from a representative normal donor in duplicate to show equal loading. GAPDH was the internal positive control, and PCR was carried out in the absence of RT product as a negative control (-). The amplicons generated were consistent with the sizes expected for GAPDH (450 bp), 5-LO (124 bp), FLAP (210 bp), LTA₄H (291 bp), LTC₄S (155 bp) and CysLT₁ (248 bp). Panel (B) shows an immunoblot of lysates from fibroblasts from a representative asthmatic patient (F) compared with peripheral blood leukocytes (PBL). Each lane represents the same amount of total cellular protein. Recombinant protein standards are shown for 5-LO and FLAP (+). n = 5 normals, n = 5 asthmatics.