Figure 5

Prolyl hydroxylase dependent regulation of Dicer. Dicer expression was examined in SKBR3 cells after inhibition of each prolyl hydroxylase by siRNAs. A, Dicer mRNA expression in SKBR3 cells after transient transfection with PHD2 targeting siRNAs or control siRNA in normoxia (P = 0.0008). *denotes P < 0.05 compared with parallel controls. Data represent normalized mean ± S.E (error bars) (n = 3). Dicer mRNA levels were analysed by RT-PCR and normalised to β-2-microglobulin mRNA levels. Statistical significance established by Student’s t-test. B, PHD2 and Dicer protein levels after transient transfection of SKBR3 cells with two independent siRNAs targeting PHD2 or control siRNAs in normoxia. Results show two technical replicates per treatment. C, Dicer, Drosha and TARBP2 protein expression after transient transfection of SKBR3 cells with siRNAs targeting PHD2 or control siRNAs in normoxia. D, Dicer protein expression after transient transfection of SKBR3 cells with siRNAs targeting PHD1 siRNAs or control siRNAs in normoxia. E, Dicer protein expression after transient transfection of SKBR3 cells with siRNAs targeting FIH-1 siRNA or control siRNA in normoxia. Results show three technical replicates per treatment. Protein expression was examined by immunoblotting. α-actinin was used as the loading control.