Smoking is significantly associated with negative affect, which may play an especially important role in the smoking behavior of women. The purpose of this laboratory study was to examine the role of gender in the relationship of negative mood and smoking maintenance for male and female smokers following an implicit mood induction using music. Ninety adult smokers (50% female) completed a laboratory session during which they were randomly assigned to a negative mood induction, a positive mood induction, or a neutral mood condition. Latency to smoke and number of cigarettes smoked were assessed during an ad libitumsmoking period following the mood induction. Female smokers began smoking more quickly following the negative mood induction when compared with males. There were no gender differences in the number of cigarettes smoked or for cravings to smoke by mood condition. This study demonstrated gender differences in the relationship between negative affect and smoking behavior following an implicit and subtle mood manipulation. A better understanding of gender differences in smoking behavior can provide valuable information about mechanisms that maintain smoking behavior and guide treatment development to help adults quit smoking.