Handwashing after using the restroom is generally poor across the population, and one common method used to increase the frequency of handwashing is to place signs reminding individuals to wash their hands. The current study examined the association between the absence and presence of signs reminding one to wash their hands and handwashing in public restrooms. Signs prompting handwashing behavior remind restroom patrons of acceptable behavior, and the presence of these signs is hypothesized to be associated with an increase in handwashing. Observation of 175 individuals (95 women and 80 men) using public restrooms on a university campus indicated that 61% of the women and 37% of the men observed washed their hands, e.g., washing hands with soap, in the absence of the sign, and 97% of the women and 35% of the men observed washed their hands in the presence of the sign. Further, 53% of the men and 38% of the women observed rinsed their hands, e.g., washing hands without soap, in the absence of the sign, and 55% of the men and 2% of the women observed rinsed their hands in the presence of the sign. Results are discussed in terms of possible factors associated with sex differences in handwashing and the absence and presence of visual prompts for handwashing behavior.