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. 2012 May 7;26(7):1144–1157. doi: 10.1210/me.2011-1343

Fig. 7.

Fig. 7.

Role of OT in maintaining ESC in an undifferentiated state. A, Real-time RT-PCR quantification of undifferentiated marker genes. Cells were treated with OTR-neutralizing antibody (0.1 μg/ml), OTR-specific siRNA, combination of OTR-neutralizing antibody and OTR-specific siRNA, CREB-, NF-κB-, CBP-, CX43-specific siRNA, 18α-GA (10 μg/ml), and ntRNA for 72 h. Total RNA from mouse ESC was reverse transcribed, and Oct4, Sox2, FoxD3, Nanog, and β-actin cDNA were amplified by real-time PCR as described in Materials and Methods. The data are reported as the mean ± se of five independent experiments, each conducted in triplicate. *, P < 0.05 vs. control. B, Cells were treated with OTR-neutralizing antibody, OTR-specific siRNA, combination of OTR-neutralizing antibody and OTR-specific siRNA, CREB-, NF-κB-, CBP-, CX43-specific siRNA, 18α-GA, and ntRNA for 72 h. Total protein was extracted and blotted with Oct-4, SSEA-1, Nanog, and β-actin antibodies. Each example shown is representative of five independent experiments. The right part of panel B depicting the bars denotes the mean ± se of five independent experiments for each condition determined from densitometry relative to β-actin. *, P < 0.05 vs. control. Con, Control; ntRNA, nontargeting RNA; ROD, Relative Optical Density.