Non-motorized travel modes, particularly cycling, are experiencing a resurgence in many United States (U.S.) states as well as in other countries. Still, most studies focus on bicyclists’ behaviors in areas with strong bicycling cultures. This paper discusses the findings of a survey (N = 1,178) deployed in six communities in Alabama and Tennessee, U.S., where cycling is not (yet) popular nor widely adopted. The analysis includes linear regression models built on respondents’ reactions to images of bicycling infrastructure and their perceptions of being comfortable, safe, and willing to try cycling on the displayed roadway type. Findings indicate a preference for more separated bicycle infrastructure types along with options that exclude on-street parking. Segmented models indicate that, compared with potential cyclists, the preferences of regular utilitarian cyclists can vary more than those of recreational/occasional cyclists. Results from this study provide useful insights into ways to maximize the return on investments, and design bike infrastructure that can attract patronage and be most successful in areas lacking a substantial bicycling population.