Gene expression profiling of all DOCK RhoGEFs in purified postnatal Purkinje neurons. (A, B) Sorting of GFP-expressing postnatal Purkinje neurons by FACS. Dissociated cells from cerebella of Pcp2-GFP mice at P3, P7, P15, and P20 were plated before (A) and after (B) sorting and fixed. Nuclei were stained with Hoechst stain to visualize the total cell number in the sample, and Purkinje neurons were revealed with a calbindin antibody (red) and by direct fluorescence of the GFP (green). Micrographs show representative sorting results of cerebellar cells from mice at P7. (C) Histogram representing the enrichment of Purkinje neurons within the cell population upon sorting, expressed as relative percentages. Data are expressed as mean enrichment value of all age groups combined ± SD of at least three independent experiments. **p < 0.01 (Student's t test). (D) RT-qPCR performed before and after FACS on the mRNA of specific marker genes of the main cerebellar cell types: calbindin (Purkinje cells), NeuroD1 (cerebellar granule neurons), GFAP (astrocytes), and Tcfap2a (interneurons). Data are expressed as mean ± SD of at least three experiments performed on P7 Pcp2-GFP mice in the example shown. **p < 0.01 and ***p < 0.001 (Student's t test). (E) RT-qPCR performed on purified PC mRNAs of all 11 mammalian DOCK-family RhoGEFs. Data are expressed as mean ± SD of at least three experiments. *p < 0.05, **p < 0.01, and ***p < 0.001 (Student's t test).