This case study concerns the characteristics of journalistic newsroom culture of a regional news desk of Czech Television, a public-service broadcaster. It seeks to explore the attributes that each of the two news desk editors promote in relation to newsroom culture and whether they are gender specific. The article also discusses whether newsroom culture constructs the same working conditions for male and female journalists. The article draws on theories developed in media and journalism studies and especially the theories and empirical findings of feminist media scholars regarding newsroom culture and the status of male and female journalists in the workplace. It presents an analysis based on qualitative data obtained during four-week participant observation in a regional newsroom of Czech Television, combined with semi-structured individual interviews about the experiences of eight selected male and female staff members working in different positions in the editorial hierarchy. This article presents the very first findings about the gendered characteristics of newsroom culture in the Czech Republic.