Fig. 1.

5-LOX mRNA expression in mouse leukocytes (L) and cerebellar granule cells (CGC). Leukocytes were extracted from whole blood as described elsewhere (Mohammadi and Saberivand, 2009). The signals for 5-LOX and cyclophin (Cyc; a loading control) mRNAs were obtained by RT-PCR using specific primers (see Table 1). The amplification step lasted for 30 cycles with denaturation (94°C, 30 sec), annealing (57°C, 30 sec), and elongation (72°C, 30 sec). Shown is an example of gel electrophoresis of the PCR products (bands at the expected size for 5-LOX and Cyc, i.e., 178 and 153 bp respectively). Note the stronger 5-LOX mRNA signal in leukocytes compared to CGC whereas the cyc signals were comparable in both samples. A real-time PCR assay of these samples revealed that leukocytes express about 400 times more 5-LOX mRNA than CGC [5-LOX mRNA (units): leukocyte = 87, CGC = 0.22; n = 2].