Re-evaluating Laminar Specificity of Working Memory in Human Prefrontal Cortex

Although working memory reliably activates the dorsolateral prefrontal cortex (dlPFC), the functional contributions of its cortical layers in humans remain unclear. A seminal study reported a laminar dissociation in dlPFC—superficial layers engaged during working-memory manipulation and deeper layers during motor response execution. Like many current layer-resolved studies, that work relied on several manual and semi-manual processing steps, including the selection of regions of interest. We conducted a preregistered replication in 21 human participants using an automated and fully reproducible analysis pipeline. While we replicated the superficial-layer effect during manipulation, we found no evidence for preferential deep-layer activation during response execution. This biologically plausible result refines current models of laminar organization in the human prefrontal cortex and aligns human evidence with the more heterogeneous animal literature. By demonstrating reproducibility through preregistration and automation, this work establishes a benchmark for laminar analyses in cognitive neuroscience.