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. 2011 Nov 22;153(1):42–55. doi: 10.1210/en.2011-1308

Fig. 4.

Fig. 4.

Real-time PCR analysis of transcript levels of (A) Nsbp1, (B) Pde4d4, and (C) Hpcal1 and cell survival of in NbE-1 and AIT cells in response to siRNA knockdown. Cells were treated with 0.2 μm siRNA oligos or with 0.2 μm nontarget control for 72 h. Left panel, RNA was isolated and reverse transcribed. Nsbp1, Pde4d4, or Hpcal1 expression was quantified by real-time PCR and normalized to Rpl19 in the same sample. The 2-ddCt method was used to calculate the relative expression level of transcript [relative expression ratio (RER)] in target-gene knockdown samples. RER of nontarget control was set as 1.0. Right panel, Percentage of viable cells (compared with nontarget control) after siRNA knockdown assay was determined by alamarBlue. No significant toxicity was found in cells treated with Lipofectamine only. Data presented are the mean with sd of four separate experiments. *, P < 0.05 or **, P < 0.01 were considered statistically significant.