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

Figure 3.

Figure 3

RNAi-mediated downregulation of MSP58 inhibited growth of U251 cells in vitro. (A) Cell growth curve. Parental U251 cells, vector control U251 cells (U251-H1 neo and U251-NC), and MSP58 shRNA-stable cells (clone U251-S and U251-Smix) were seeded into 96-well tissue culture plates. Cell viability was measured using the MTT assay. Cell growth curves were determined by reading the absorbance at 490 nm on a microreader. (B) Cell cycle distribution. Parental U251 cells, vector control U251 cells (U251-H1 neo and U251-NC), and MSP58 shRNA-stable cells (clone U251-S and U251-Smix) were stained with propidium iodide. The DNA content of the cells was analyzed by FACS. (C) Cell apoptosis in shRNA stably transfected U251 cells. Vector control U251 cells (U251-NC) and MSP58 shRNA-stable clones (U251-S, U251-Smix) were stained by AnnexinV/propidium iodide and analyzed by FACS to detect apoptosis. U87 cells treated by cisplatin (5 μg/ml, 48h) were used as positive control. (D) Cell apoptosis in siRNA-treated U251 cells. U251 cells were transfected transiently with non-silencing control siRNA or siRNA targeting MSP58. 48 hours later, apoptosis was analyzed by FACS. Values are mean ± SD of three independent experiments. *P < 0.05 versus U251, U251-H1 neo, and U251-NC control cells.