Figure 2

Analysis of lamin knock-down in U-2 OS cells seeded on an adenoviral microarray. A. Fluorescent micrographs of cells expressing RFP and shCTR stained with antibody directed against lamin A/C. Red fluorescence (left) indicates infected cells, green fluorescence (right) indicates lamin A/C expression. The diagram illustrates the evaluation procedure. Scale bar, 10 μm. B. Bar diagram, showing the averages of fluorescence intensity of the lamin A/C immunostaining of U-2 OS cells expressing either an irrelevant shRNA (shCTR), or shRNA directed against lamin A/C (shLamin) [5] for the experiments performed on the chip (black bars) or in the 96-well plate (gray bars). Infection efficiency achieved on the microarray as well as in the well suffices to elicit a significant knock-down (4 spots, n > 150, p < 0.005; by ANOVA analysis). C. Validation of shRNA targeting Lamin A/C by Western immunoblot analysis. U-2 OS cells were transfected either with pENTR +pcDNA3-eGFPneo (lane 1) or with pENTR-shLaminA/C +pcDNA3-eGFPneo (lane 2) and the lysates were immunoassayed 48 hours later.