Background Stair climbing helps to accumulate short bouts of physical activity throughout the day as a strategy for attaining recommended physical activity levels. There exists a need for effective long-term stair-climbing interventions that can be transferred to various worksite settings. The aims of this study were: 1) to evaluate short- and long-term effectiveness of a worksite stair-climbing intervention using an objective measurement of stair climbing and a controlled design; and 2) to perform a process evaluation of the intervention. Methods We performed a controlled before-and-after study. The study was conducted in two corporate buildings of the same company located in Paris (France), between September, 2013 and September, 2014. The status of either “intervention site” or “control site” was assigned by the investigators. Participants were on-site employees (intervention site: n = 783; control site: n = 545 at baseline). Two one-month intervention phases using signs (intervention phase 1) and enhancement of stairwell aesthetics (intervention phase 2) were performed. The main outcome was the change in stair climbing, measured with automatic counters and expressed in absolute counts/day/100 employees and percent change compared to baseline. Qualitative outcomes were used to describe the intervention process. Results Stair climbing significantly increased at the intervention site (+18.7 %) but decreased at the control site (-13.3 %) during the second intervention phase (difference between sites: +4.6 counts/day/100 employees, p < 0.001). After the intervention and over the long term, stair climbing returned to baseline levels at the intervention site, but a significant difference between sites was found (intervention site vs. control site: +2.9 counts/day/100 employees, p < 0.05). Some important facets of the intervention were implemented as intended but other aspects had to be adapted. The main difficulty reported by the company’s staff members lay in matching the internal communications rules with critical intervention criteria. The program was maintained at the setting level after the end of the study. Conclusions This study shows a successful stair-climbing intervention at the worksite. The main barriers to adoption and implementation were related to location and visibility of posters. Process evaluation was useful in identifying these barriers throughout the study, and in finding appropriate solutions.