Fig. 2

Mark4 up-regulation after subcortical stroke in Layer 5 cortical neurons. Image of fluororuby+ labeling in ipsilateral cortex after stroke (arrow) in sections prepared for laser capture microdissection (left, a). Individual fluororuby+ cortical neurons before (upper panel) and after (lower panel) laser capture microdissection (right, a). Graph of qPCR fold expression differences normalized to neurofilament heavy chain (NFH) for cell-type specific marker genes from laser-capture RNA (red) compared to whole brain RNA (black) (b) (p < 0.0001 by one-way ANOVA with adjusted p-value for Fezf2 levels < 0.0001 by Sidak’s correction). Agarose gel of Mark4 PCR product from pooled LCM isolates (lower panel, c). qPCR for Mark4 in laser-captured FR- (L5) or FR+ (FR + L5) Layer 5 cortical neurons (1.3-fold increased expression, confidence interval ± 0.18, n = 4 in L5, n = 8 in FR + L5) (upper panel, c). Mark4 protein expression (green) in fluororuby+ (red) stroke-injured cortical neurons 7 days after stroke (d). Number of Mark4+ cells (bar plots with quartiles) and average intensity of Mark4 expression (AFUs/unit area) in FR- and FR+ cortical neurons after stroke (e) (n = 12, total cells = 240, p = 0.0053 for average Mark4 AFUs per animal by t-test). Mean ± S.E.M. Scale bars = 10 μm