Small interfering rna a as control sc 37007

The RNAPII inhibitor, α-amanitin (A2263 Sigma, St Louis, MO, USA) was dissolved in bi-distilled sterile water and filtered by 0.2 μm filter. The experiments were performed by using growing doses of a-amanitin, that is, 1, 2, 3, 5, 10 μg/ml on each point of 2000.000 cells/ml or 2 and 10 μg/ml on each point of 500 000–1000 000 cells/ml. After 16 h, the cells were lysed in 700 μl of Qiazol (Qiagen, Venlo, Netherlands) for the RNA extraction.
Transfection of DNA vectors (200–500 ng/50 000 cells) in HEK293T was performed with Lipofectamine2000 (Invitrogen, Carlsbad, CA, USA) in accordance with the manufacturer's procedures, whereas transfection of MEC-1, MEC-2, WaC3CD5 and K562 was performed using the Amaxa nucleofector (Lonza, Basel, Switzerland) according to the manufacturer's protocols (plasmids:3000 ng/1 000 000 cells; small interfering RNA 40 pmol/1 000 000 cells). siPOLR3A (sc-90684), siPOLR3G (sc-43507) and small interfering RNA -A as control (sc-37007) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The efficiency of transfection was determined by cytofluorometry, messenger RNA expression and\or fluorescence microscopy. After transfection of luciferase reporter vectors (pGL3Ren-187_A and pGL3Ren-187_G), the luciferase assay was conducted according to manufacturer's dual luciferase assay protocol (Promega).