Figure 9

Knockdown of h NAA16 downregulates hNaa10p. (A) HeLa cells cultured in 6 cm dishes were transfected with 50 nM unspecific (Ctr), hNAA15 or hNAA16 siRNAs. ZVAD-fmk (5 μM) was added 24 hours post transfection to prevent any induction of apoptosis. After 48 hours total RNA was isolated and processed by RT-PCR with specific primers to hNAA15, hNAA16 and β-actin. (B) HeLa cells cultured in 6 cm dishes were transfected with 50 nM unspecific (Ctr), hNAA15, hNAA16 or hNAA10 siRNAs. ZVAD-fmk (5 μM) was added 24 and 48 hours post transfection to prevent any induction of apoptosis. After 72 hours cell lysates were analyzed by SDS-PAGE/Western blotting. The membrane was incubated with anti-hNaa10p, anti-hNaa15p and anti-β-tubulin. Results shown are representative of three independent experiments. Protein levels were quantified using FUJIFILM IR LAS 1000 and Image Gauge 3.45. Protein levels in siCtr (-) samples were set to 1.0 and protein levels in sihNAA15, sihNAA16 or sihNAA10 treated cells were estimated relative to this and normalized to β-tubulin levels.