The gender-based stereotype of nursing as a female profession has been a large obstacle to men entering the nursing profession. However, there is little quantitative research on the influence of prejudice induced by this stereotype on male nursing students' willingness to be nurses. To examine the effect of perceived prejudice on willingness to be a nurse via the mediating effect of satisfaction with major among Chinese male nursing students. A cross-sectional survey was used. Four hundred and sixty male nursing students who were enrolled either in bachelor's programs in universities or advanced diploma programs in colleges in Jinan, China, were surveyed using questionnaires measuring perceived prejudice, satisfaction with major, and willingness to be a nurse. Structural equation modeling with bias-corrected bootstrapping was employed to determine the influence of perceived prejudice on willingness to be a nurse with major satisfaction as a mediator. Male students who were in an advanced diploma nursing program and those for whom nursing was the first-choice major reported significantly less perceived prejudice, greater satisfaction with major, and greater willingness to be nurses than did those in a bachelor's nursing program and those for whom nursing was not the first-choice major, respectively. Moreover, although perceived prejudice had no significant direct effect on willingness to be a nurse (β=0.07, p>0.05), it did have a strong indirect effect (full mediation) via satisfaction with major (β=−0.59, p<0.001). Perceived prejudice strongly influenced male nursing students' willingness to be nurses via the full mediating effect of satisfaction with major. Because this obsolete stereotype of nursing as a female occupation gives birth to prejudice against male nursing students, effective measures should be taken to change this stereotype to recruit more men as nursing staff. •Perceived prejudice had a strong indirect effect on willingness to be a nurse.•Perceived prejudice had no significant direct effect on willingness to be a nurse.•Measures should be taken to change the stereotype of nursing as women's profession.