The aim of the present study was to investigate the effect of cultural knowledge on improving Iranian EFL learners' listening comprehension. To achieve this purpose, out of 300 participants, 120 pre-intermediate language learners were selected based on their scores on a listening comprehension test and were randomly assigned to four groups. Each group was exposed to a certain condition as follows: TC (Target Culture), ITC (International Target Culture), SC (Source Culture), and CF (Culture Free). At the end of the experiment, to see whether or not any changes happened regarding their listening proficiency, a post-test was administered to the four groups. The results suggested that the participants performed differently on the post-test indicating that familiarity with culturally-oriented language material promotes the Iranian EFL learners' listening proficiency. (Contains 7 tables.)