FIGURE 1.

(A) Schematic of snoRNA knockdown in human cultured cells. The chimeric ASO (αU84) targeting U84 snoRNA is shown. The five terminal nucleotides on each end are 2′-O-methoxyethyl nucleotides indicated as “mN,” where N is the nucleotide. The phosphothioate backbones are indicated with asterisks. Oligonucleotides were introduced into human cultured cells using a nucleofector device. Total RNA was prepared and used for RPA. (B) Oligonucleotide dose-dependency in the knockdown of three snoRNAs (U84, HBII295, and H/ACA38) as measured by RPA. Numbers shown above the blot represent oligonucleotide amounts (pmol) used for nucleofection. Pr and Y represent the RNA probe and control yeast RNA used for RPA, respectively. (*) Putative degradation products. (C) Effects of ASOs and siRNAs. ASOs and siRNAs with complementarity to GFP (αGFP) or U84 (αU84) were administered by nucleofection. Total RNA was prepared at 6 or 24 h after nucleofection (as shown above the panel). (D,E) Time course of knockdown efficacy of αU84 (D) and αHBII295 and αH/ACA38 (E). Numbers above the blots represent incubation time (hours) after nucleofection. U84 levels are shown after oligonucleotide administration with the lipofectin reagent (three right lanes in D).