For decades, the importance of highway work zone safety has increased considerably with the continual increase in the number of highway work zones present on highways for repairs and expansion. Rural work zones on two-lane highways are particularly hazardous and cause a significant safety concern due to the disruption of regular traffic flow. In this study, researchers determined motorists’ responses to warning signs in rural, two-lane highway work zones. The researchers divided vehicles into three classes (passenger car, truck, and semitrailer) and compared the mean change in speed of these classes based on three different sign setups: portable changeable message sign (PCMS) OFF, PCMS ON with the message of Slow Down, Drive Safely, and a temporary traffic sign (W20-1, “Road Work Ahead”). Field experiments were conducted on two two-lane work zones with flagger control. Statistical analyses were performed to determine whether there was a significant interaction between motorists’ responses and the sign setups. Data analysis results show that a visible PCMS, either turned on or off, was most effective in reducing truck speeds in rural, two-lane work zones. The temporary traffic sign (W20-1) was more effective in reducing the vehicle speeds of passenger car and semitrailer. Results of this research project will help traffic engineers to better design the two-lane work zone setup and take necessary safety countermeasures to prevent vehicle crashes.