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. 2004 Jul 12;32(12):e100. doi: 10.1093/nar/gnh098

Figure 7.

Figure 7

Ectopic expression of hMLH1 increased MMR efficiency in HCT116 cells. (A) Detection of hMLH1 protein with western blotting. HCT116 and SW480 were co-transfected with 1 μg pCMV-MLH1 or 0.6 μg pRC/CMV (control) and the homo- or heteroduplex (with a G–G mismatch) plasmid. Forty-eight hours later, the transfected cells were lifted and analyzed with flow cytometry. Cells recovered from the flow cytometer, along with HCT116+ch3 and SW480 cells, were lysed. Proteins were extracted, separated on 8% SDS-PAGE, and immunoblotted by a hMLH1 antibody. (B) Effect of hMLH1 transfection on G–G MMR in HCT116 and SW480 cells. Relative EGFP expression was calculated after flow cytometry analysis as described in the Materials and Methods. According to student t-test, HCT116 transfected with hMLH1 expression plasmid has a significant higher (P < 0.02) MMR efficiency (0.43 ± 0.05) than HCT116 transfected with pRC/CMV (0.26 ± 0.05). Each column represents mean ± SEM of three independent measurements.