Table 1.
siRNA primers
| Gene name | Primer |
|---|---|
| Rpn11 | 5′-CAAGAGCUUUCUUUAAGAA-3′ (positions 230-248 of human mRNA) |
| 5′-GUUGUACCUGCCAGAAUUA-3′ (positions 210-228 of human mRNA) | |
| UCHL5 (Uch37) | 5′-CUUAGAGCAACAUCCUAAU-3′ (positions 1192-1211 of human mRNA) |
| 5′-GAGAAGGACCGAUUGAUUU-3′ (positions 676-694 of human mRNA) | |
| Usp14 | 5′-AGAAAUGCCUUGUAUAUCA-3′ (positions 1135-1153 of human mRNA) |
| 5′-GCAUUGCAGGGAAACUUAU-3′ (positions 3406-3425 of human mRNA) | |
| S4 | 5′-CAUGCACCGUCCAUCGUGU-3′ (positions 877-895 of human mRNA) |
| b5 | 5′-GAAGGUGAUAGAGAUCAAC-3′ (positions 292-310 of human mRNA) |
| EGFP | 5′-GGUGAACUUCAAGAUCCGC-3′ (positions 654-672 of human mRNA) |
RT-PCR. Total RNA was isolated from HeLa cells using Trizol reagent (Invitrogen) according to the manufacturer's instructions. RNA content was determined by measuring A260. RT-PCR was carried out using OneStep RT-PCR kit (Qiagen) using pairs of primers amplifying the genes of interest, as indicated in Table 2. The number of cycles was adjusted to obtain a linear range of reaction products.