fig6

Figure 6. (A) ERK expression in T24 cells 72 h after MMC treatment alone. ERK staining showed progressive decline in the staining intensity with increasing MMC dose; (B) ERK staining with MMC combined with siRNA, where the decline in staining intensity was greater when MMC was combined with ras siRNA. In all slides, the apoptotic and mitotic cells (arrows) were negative for ERK staining (200×). To the right, overlay histograms of flow cytometry show the extent of ERK specific fluorescence (red-filled) in comparison to isotype control fluorescence (unfilled). Western blot traces show mild decrease of the blot thickness and intensity at 10, 30, and 100 μg/mL MMC compared to the UT control (above), with nearly complete absence of the blot when MMC was combined with ras siRNA (below). All the bands appeared between 39 and 45 kDa on the molecular marker at the standard ERK 1/2 molecular weight of 42/44 kDa. MMC: mitomycin C; UT: untreated cells