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It is generally believed that 5αR2, rather than 5αR1, plays a key role in the pathogenesis of male-pattern baldness. Next, we determined 5αR2 mRNA RG108 levels in the three cell types using real-time RT-PCR analysis. Barely detectable levels of 5αR2 mRNA were observed in the three cell types (Fig. 2B). No significant difference in 5αR2 mRNA expression levels was observed.
Protein expression and cellular distribution of 5αR
To further identify levels of 5αR protein in cultured DPCs and dermal fibroblasts, Western blot analysis was performed after total cellular protein was extracted from the three cell types after four passages. A faint immunoreactive band at 26 kDa corresponding to the protein for human 5αR1 was detected in either bDPCs or sDPCs, but a similar band was not detected in dermal fibroblasts (Fig. 3A). Densitometric analysis demonstrated that the level of 5αR1 expression in bDPCs was approximately 1.6-fold higher than that in sDPCs by using β-actin as an internal housekeeping marker to normalize the signals obtained for 5αR1 in each experiment (Fig. 3B).