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Figure 1 | Molecular Cancer

Figure 1

From: Global gene expression profiling of cells overexpressing SMC3

Figure 1

Identification and characterization of 293 cells stably overexpressing human SMC3. a): 293 cells were transfected with SMC3-pcDNA3.1 expression vector or the empty vector alone. Cell clones with stably integrated vector were selected in medium containing 500 μg/ml of G418 for three weeks. The expression of SMC3 and of the housekeeping gene G3DPH in drug-resistant clones was evaluated by semi-quantitative RT-PCR. b): SMC3 expression analysis was repeated on cells at later passages to confirm that clones stably overexpressing SMC3 were used. This time SMC3 expression was examined by Western immunoblotting. For this purpose cells were solubilized in lysis buffer and analyzed by 8% SDS-PAGE. After transfer of the proteins to nitrocellulose, SMC3 was immunodetected using a goat polyclonal antibody followed by ECL detection of the immunocomplex. The expression of α-tubulin was used to assess the inter-sample variability using nitrocellulose filters stripped of the SMC3 immunocomplexes.

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