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Figure 2. Expression of semaphorin, neuropilin, and plexin transcripts. (A) Reverse transcriptase polymerase chain-reaction (RT-PCR) analysis of transcripts expressed by gingival fibroblasts (G), dermal fibroblasts (D), periodontal ligament fibroblasts (P), and pre-osteoblasts (O). Relative RNA transcript levels for the semaphorins, neuropilins, and plexins were compared. The expression of the ß1 subunit of integrin acts as a loading control, since it is expressed uniformly by all of the cells examined. The photo represents a typical result from 1 of 3 experiments. RT-PCR was performed for 25 cycles in this experiment. All RT-PCR reactions were repeated with 30 cycles for those transcripts that were initially detected poorly at 25 cycles. The PDL and gingival fibroblasts represented in panel (A) were isolated from the same individual. The expression of each transcript was compared between fibroblast populations (B) and fibroblast and pre-osteoblast populations (C). Four independent isolates of dermal, PDL, and gingival fibroblasts were compared, and the comparison of expression was represented as the ratio of the mean transcript expression. Error bars represent the standard deviation. In general, a five-fold difference in expression was determined to be statistically significant by ANOVA analysis. (D) Those semaphorins and plexins with a greater-than-five-fold difference in expression are listed.
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