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. 1999 Feb;19(2):1101–1115. doi: 10.1128/mcb.19.2.1101

FIG. 2.

FIG. 2

Regulation of cyclin D1 and c-Myc expression by Raf. (A) Cycloheximide sensitivity of Raf-induced cyclin D1 and c-Myc mRNA expression. NIH 3T3 cells expressing ΔB-Raf:ER* were cultured in 0.5% FCS for 24 h and were then either left untreated or treated with 25 μg of cycloheximide per ml for 1 h. These cells were then either left untreated (CHX) or treated with 1 μM ICI to activate ΔB-Raf:ER* for different lengths of time as indicated. The expression of cyclin D1, c-Myc, and GAPDH mRNAs were quantitated by RNase protection assays. (B) Cyclin D1 promoter activation. NIH 3T3 cells expressing ΔB-Raf:ER* were transiently transfected with reporter constructs consisting of the promoter region of mouse cyclin D1 linked to luciferase [mD1(-984)Luc] and pSRαβ-Gal as a control for transfection efficiency. Transfected cells were treated with either ethanol (solvent control, open bar) or 1 μM 4-HT (closed bar) for 41 h, at which time luciferase and β-galactosidase activities were measured. Results are presented as the ratios of the luciferase to the β-galactosidase activities.