This study investigated differentiated instruction strategies presented in pre-service EFL teachers’ teaching modules, the challenges they encountered, and their coping strategies. Using qualitative case study design, five pre-service EFL teachers enrolled in a teacher professional development program were involved as participants. To answer the objectives of this study, document analysis of the teaching modules and interview transcripts was analyzed using data condensation, data display, and conclusion drawing techniques. The findings revealed that the pre-service EFL teachers applied various strategies across three differentiation domains. In content differentiation, they involved pre-assessment, student-related considerations, delivery modes, and teaching media. Moreover, process differentiation was frequently evident, including student grouping, classroom activities, assignment formats, scaffolding, and technology integration. Therefore, product differentiation was limited to the presentation formats of products. Furthermore, temporal differentiation practice was also identified in post-learning activities. On the other hand, the teachers’ challenges included limited understanding, grouping issues, limited resources, and time constraints. To address these challenges, they involved in professional collaboration, modified instruction, resource utilization, and time management to cope with their struggles. Consequently, this study highlights the need for structured support and practical training on differentiated instruction, promoting inclusive and responsive teaching practices in teacher education.