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. 2006 Jan 19;25(3):522–532. doi: 10.1038/sj.emboj.7600942

Figure 7.

Figure 7

PACT is required for the accumulation of mature miRNA. (A) Northern blot analysis. siRNA targeting Dicer, TRBP, or PACT was transfected into a HeLa cell line that expresses primary miR-30a under the control of the tetracycline-inducible promoter. Following transfection (48 h), doxycycline (3 μg/ml) was added to the medium to induce the expression of miR-30a. After 24 h, the cells were harvested for Northern blotting (the left panel). After normalization for loading, the relative ratios for mature miR-30a were calculated and normalized to no doxycycline-treated experiment (lane 1). The average value obtained from two independent experiments was presented in the right panel. (B) The same experiment as described in (A), except for using pri-miR-124a inducible HeLa cell line. The left panel shows a representative Northern blot result. The right panel presents the mean value of relative expression level for mature miR-124a obtained from two independent experiments. (C) RT–PCR. Total RNA used in (A) was reverse-transcribed and amplified by PCR using gene-specific primers.