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Using TALIS 2018 U.S. data, this study investigated the direct and indirect effects of teacher self-efficacy in multicultural classrooms (TSMC), professional development in multicultural education (PD in ME), and the percentage of English Learner (EL) students on STEM teachers’ instructional practices. Path analysis revealed that PD in ME significantly enhanced STEM TSMC, while the percentage of EL students showed no significant impact. Increased TSMC significantly improved clarity of instruction, classroom management, and cognitive activation. Indirectly, PD in ME significantly boosted instructional practices through increased TSMC, whereas the percentage of EL students did not have a significant indirect effect. These findings highlight the importance of specialized interventions to enhance TSMC and improve STEM teachers’ instructional practices in diverse environments.