The paper investigates gender biases and differential treatment of women and men in the business start-up phase. A sample of 498 entrepreneurs from Slovakia participated in an online experiment and evaluated three fictitious business plans in terms of the applicants’ competence, likeability, and business ability. The start-ups were positioned in three different sectors—cosmetics production, services provision, and software development—where men’s and women’s chances of success may be viewed differently. Following Goldberg’s paradigm, half of the evaluators received business plans presented as written by female and half by male applicants; otherwise the plans were identical. Results imply that female applicants are assessed similarly to male applicants, but more masculine evaluators assess women’s business plans and their potential in entrepreneurship more critically. The study advises caution in recommending more female evaluators in the business plan assessment. If women who become involved in entrepreneurship are excessively masculine and masculinity is associated with a less favourable evaluation of potential female entrepreneurs, such policies could backfire against women, putting them in a more disadvantaged position.