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This study aimed to examine the associations between participation frequency in physical education (PE) classes and objective measures of physical activity (PA) and sedentary behavior (SB) in children from 12 countries at different levels of development.This multinational, cross-sectional study included 5874 children 9–11 yr old from sites in Australia, Brazil, Canada, China, Colombia, Finland, India, Kenya, Portugal, South Africa, the United Kingdom, and the United States. PA and SB were monitored for seven consecutive days using a waist-worn accelerometer. PA and SB data were presented for weekdays (times in and out of school) and weekend days. Participation frequency in PE classes was determined by questionnaire. Multilevel modeling analyses stratified by sex were used.Overall, 24.8% of children self-reported participation in PE classes ≥3 times per week (25.3% in high-income countries [HIC] and 24.3% in low- and middle-income countries [LMIC]). After adjusting for age, sex, parental education, and body mass index z-score, results showed that children from low- and middle-income countries who took PE classes one to two times per week were more likely to present better indicators of PA and shorter time in SB in and out of school. In HIC, boys that participated in PE classes were more likely to meet the moderate-to-vigorous PA recommendations and to present better indicators of PA (in school) and shorter time in SB in and out of school. For girls in HIC, attending PE classes increased the likelihood of spending more time in moderate-to-vigorous PA, especially if they attended ≥3 times per week.Attending PE classes is associated with a higher level of PA and a lower level of SB in and out of school during weekdays in children from countries at various levels of development.