Gender inequity in science and academia, especially in senior positions, is a recognised problem. The reasons are poorly understood, but include the persistence of historical gender ratios, discrimination and other factors, including gender-based behavioural differences. We studied participation in a professional context by observing question-asking behaviour at a large international conference with a clear equality code of conduct that prohibited any form of discrimination. Accounting for audience gender ratio, male attendees asked 1.8 questions for each question asked by a female attendee. Amongst only younger researchers, male attendees also asked 1.8 questions per female question, suggesting the pattern cannot be attributed to the temporary problem of demographic inertia. We link our findings to the 'chilly' climate for women in STEM, including wider experiences of discrimination likely encountered by women throughout their education and careers. We call for a broader and coordinated approach to understanding and addressing the barriers to women and other under-represented groups. We encourage the scientific community to recognise the context in which these gender differences occur, and evaluate and develop methods to support full participation from all attendees.