PSD-95 interacts with EB3. A, Colocalization of EB3 comets with PSD-95 puncta. Primary cultures of hippocampal neurons were fixed with ice-cold methanol followed by 4% paraformaldehyde. Double-label immunocytochemistry with antibodies to PSD-95 (left) and EB3 (middle) was performed. Images are overlaid for direct comparison (right). Inset, High magnification indicating colocalization of PSD-95 and EB3 puncta (arrows). B, Colocalization of EB3 and PSD-95 peaks at DIV12. Colocalization was quantified by calculating the proportion of total EB3 comets that overlay PSD-95 puncta. ***p < 0.001 by ANOVA followed by a Bonferroni multiple comparisons test. C, Coimmunoprecipitation of EB3 with PSD-95 from adult rat brain. EB3 coimmunoprecipitates with PSD-95. Detergent extracts from adult rat brain were subjected to immunoprecipitation with the indicated antibodies. IP, Immunoprecipitation; WB, Western blot.