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Abstract Background Cross-sectional studies have reported associations between social support and health, but prospective evidence is less conclusive. This study aims to investigate the associations of positive and negative experiences of social support with current and future lifestyle factors, biological risk factors, self-perceived health and mental health over a 10-year period. Methods Data were from 4,724 Dutch men and women aged 26-65 years who participated in the second (1993-1997) and in the third (1998-2002) or fourth (2003-2007) study round of the Doetinchem Cohort Study. Social support was measured at round two using the Social Experiences Checklist. Health was assessed by several indicators such as smoking, alcohol consumption, physical activity, fruit and vegetable intake, overweight, hypertension, hypercholesterolemia, self-perceived health and mental health. Tertiles of positive and negative experiences of social support were analysed in association with repeated measurements of prevalence and incidence of several health indicators using generalised estimating equations (GEE). Results Positive and negative experiences of social support were associated with prevalence and incidence of poor mental health. For the lowest tertile of positive support, odds ratios were 2.74 (95% CI 2.32-3.23) for prevalent poor mental health and 1.86 (95% CI 1.39-2.49) for incident poor mental health. For the highest tertile of negatively experienced support, odds ratios for prevalent and incident poor mental health were 3.28 (95% CI 2.78-3.87) and 1.60 (95% CI 1.21-2.12), respectively. Low levels of positive experiences of social support were also associated with low current intake of fruits and vegetables, but not with future intake. Negative experiences of social support were additionally associated with current smoking, physical inactivity, overweight and poor self-perceived health. Furthermore, high levels of negative experiences of social support were associated with future excessive alcohol consumption (OR 1.42; 95% CI 1.10-1.84), physical inactivity (95% CI 1.28; 1.03-1.58) and poor self-perceived health (OR 1.36; 95% CI 1.01-1.82). Conclusions This study showed that social support might have a beneficial effect on lifestyle and health, with negative experiences of social support affecting lifestyle and health differently from positive experiences of social support.