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. 2008 Sep 16;13(11-12):4608–4622. doi: 10.1111/j.1582-4934.2008.00499.x

Figure 2.

Figure 2

RNA interference reduced expression of MSP58 in U251 cells. (A) Down-regulation of MSP58 protein expression in clone U251-S1, S2 and S compared with control-transfected clone. (B) Down-regulation of MSP58 protein expression in clone U251-Smix. Clone U251-S1, S2 and S were pooled together to produce a mixture clone U251-Smix. (C) Down-regulation of MSP58 mRNA expression in clone U251-S was confirmed by semi-quantitative RT-PCR (C) and Western blot (E). (D) MSP58 was knocked down by transient transfection with the synthetic MSP58-targeting siRNA oligonucleotides. The expression of MSP58 in siRNA-treated U251 cells was confirmed by semi-quantitative RT-PCR (D) and Western blot (F). GAPDH was also amplified as an internal control for the RT-PCR, and β-actin levels were examined to show that similar amounts of cell lysate used in the Western Blot. U251: the parental cells; U251-H1 neo: U251 cells transfected with empty vector pSilencer3.1-H1 neo; U251-NC: U251 cells transfected with vector pSilencer3.1-NC; U251-S, S1, S2 and Smix: different U251 clones transfected with pSilencer3.1-MSP58. siRNA-NC: the transient transfectants with a negative siRNA duplex; siRNA-MSP58: the transient transfectants with a MSP58 specific siRNA duplex.