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. 2007 Oct 29;28(1):511–527. doi: 10.1128/MCB.00800-07

FIG. 6.

FIG. 6.

At a constant level of ERK2 activity, ERK1 silencing further slows cell proliferation. NIH 3T3 cells in a 10-cm dish were transfected with a total of 100 μg of plasmid containing 3 μg of pBabePuro plasmid for selection and 97 μg control plasmid (lane 1), 27 μg of pSUPER-ERK2 plasmid and 70 μg of control plasmid (lane 2), 57 μg of pSUPER-ERK2 plasmid and 40 μg of control plasmid (lane 3), or 57 μg of pSUPER-ERK2 plasmid and 40 μg of pSUPER-ERK1 plasmid (lane 4). After selection, cells were plated under conditions of exponential growth. (A) The levels of ERKs and phosphorylated ERKs were evaluated by immunoblotting at 2.5 days postplating as described for Fig. 2. The level of actin was determined for loading normalization. Results of a blot of a 14% SDS-polyacrylamide gel, which minimizes size differences between ERK1 and ERK2, for quantification by light capture are shown. The level of actin is given for loading normalization. (B) The cell proliferation assay and data presentation are as described for Fig. 2. These data are representative of three similar experiments.