Expression of E-box and ID protein mRNAs during Sertoli cell development in freshly isolated cells. The mRNAs isolated from 5- and 11-day-old Sertoli cells were analyzed by qRT-PCR using primers for Usf1 (A), Usf2 (B), Tcfe2a (E47) (C), Trf (D), and Id1, Id2, Id3, and Id4 (E–H). Also analyzed were PCR reactions performed on RT reactions lacking RT (−RT) and PCR reactions performed in the absence of cDNA (no DNA) (data not shown). Data were analyzed using the ΔΔCt method, and quantitation of the mean (±SEM) of three individual experiments for each condition is provided for each primer set. The relative mRNA levels were normalized to Ppia levels and were made relative to 5-day-old Sertoli cells (=1). Statistically significant differences (P < 0.05) are indicated by an asterisk (*).