Expression of mRNA transcripts for NGF, BDNF, and NT-3 in human brain, HMC-1 cells, and human umbilical cord blood-derived mast cells (99% purity according to staining for tryptase). Poly A+ mRNA (human brain) or total RNA treated with DNase I (for both HMC-1 cells and human umbilical cord blood-derived mast cells) and heparinase I (for human umbilical cord blood-derived mast cells only) was reverse transcribed. The cDNAs were amplified using specific human NGF primers (T and U), BDNF primers (V and W), and NT-3 primers (X and Y). As a negative control for genomic DNA contamination, DNase I-treated RNAs from HMC-1 cells and from human umbilical cord blood-derived mast cells were amplified by PCR without the reverse transcription procedure (as shown in lanes labeled as “no RT”). The resulting PCR products were analyzed by Southern hybridization using 32P-labeled cDNA probes.
