Abstract This study examined for differences in subjective ratings of discomfort and comfort (Numerical Rating Scale) and objective measures of hamstring (Sit-Reach test), lumbar (Schöber's test) and neck (Cervical Range of Motion) flexibility in healthy young subjects (n=24) following 4h of sitting on stacking chairs with or without limited legroom. When comparing the limited and unlimited legroom groups for differences in subjective and objective measures over 4h, no significant findings were seen at the 5% level of confidence. However, differences in buttock, neck, shoulder and average discomfort were significantly negatively correlated to differences between post-warm up Sit-Reach scores (Correlation Co-efficients:−.763,−.434,−.408, and−.445; p values of .004, .034, .048, and .029, respectively). The difference in buttock discomfort was significantly negatively correlated to the difference between pre-warm up Sit-Reach scores (Correlation Coefficient:−.750; p=0.005), post-sitting/pre-warm up and pre-sitting/post-warm up Sit-Reach scores (Correlation Coefficient:−.756; p=0.004), and Schöber's tests (Correlation Coefficient: −.578; p=0.049). Although the above results suggest a relationship between a loss in flexibility and an increase in discomfort, the mechanism influencing this relationship is not clear from this study. What does appear clear is that the limiting of legroom to the parameters used in this study does not seem to exacerbate change in flexibility and discomfort which are a consequence of prolonged sitting.