Figure 1.
siRNA to hKSR-2 selectively reduces the levels of the cognate mRNA and protein. A, HL60-G cells were transfected with either control siRNA (Co-siRNA), siKSR1, or si-hKSR-2 (all 5 μmol/L), incubated for 48 h, and treated with 10 nmol/L 1,25-(OH)2D3 for another 24 h. mRNA levels of KSR1 and hKSR-2 were determined by semiquantitative reverse transcription-PCR. Note that both KSR1 and si-hKSR-2 mRNA levels were induced by 1,25-(OH)2D3 treatment, and siKSR-1 or si-hKSR-2 selectively reduced the cognate mRNA expression. β-Actin RNA was used as a housekeeping gene control. The ratios of absorbance of each gene to β-actin are shown below each lane. B, protein levels of hKSR-2 in cells transfected with si-hKSR-2. Crk-L was used as a loading control. The experiments were repeated three times with similar results.