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. Author manuscript; available in PMC: 2009 Mar 5.
Published in final edited form as: J Biol Chem. 2004 Apr 15;279(27):28209–28219. doi: 10.1074/jbc.M403553200

Fig. 9. NPM siRNA inhibits MnSOD gene expression.

Fig. 9

HepG2 cells were transfected with control RNA or siRNA (double-stranded siRNA in Ready pSIREN-Shuttle vector) along with MnSOD promoter-and enhancer-driven luciferase reporter vectors for 24 h. Cells were then washed with 1x PBS and incubated in medium at 37 °C. After 12 h of co-transfection, cells were treated with TNF-α (200 units/ml) for 12 h, and then cell lysates were collected and luciferase activity was measured. A, control RNA or NPM siRNA transfected into HepG2 cells was used to isolate total RNA or total cell lysates. B, RT-PCR of RNA isolated from control and TNF-α (200 units/ml for 12 h) -treated cells was carried out using primers for each specific gene as described under “Experimental Procedures.” C, proteins from total cellular extracts were analyzed by 10% SDS-PAGE, transferred onto nitrocellulose membrane, and probed with MnSOD antibody. The same membrane was re-probed with NPM antibody (B23) or β-actin antibody. Statistical analyses were performed in three independent experiments. **, p < 0.01 compared with the corresponding control; #, p < 0.01 compared with the corresponding treatment.