Figure 1
Rational RNAi screen identifies new targets in MPM. (A) Based on the previous reports, 40 genes were selected for knockdown screening. Growth inhibition was measured in four MPM cell lines. Black boxes represent >50% inhibition with two independent siRNAs for specified target. mRNA expression following siRNA transfection was measured for each siRNA in H28 cells—black represents >75% knockdown for both siRNA; grey is >75% for one siRNA only. (B) Cells were transfected with 0.2 nM (♦), 1 nM (▴) and 5 nM (▪) siRNA targeting CDK1, PLK1 or NDC80. Cells were harvested at the indicated times and the effect on cell growth measured. Data were compared with cells transfected with a control non-silencing siRNA [C81] (○). Values are expressed as mean±s.d. (C) Cells were transfected with 1 nM siRNA in 96-well plates, and 24 h later were transferred to 6-well plates. After incubation for a further 10–14 days, cell was fixed and stained with crystal violet. (D) Protein levels in MSTO-211H and H28 cells treated with 1 nM siRNA were determined by western immunoblotting. Representative pictures are shown. β-Actin acted as a loading control. Similar results were obtained from all other cell lines (data not shown).
