Differential gene expression analysis to identify canonical Wnt signaling target genes in QR neuroblast descendants in C. elegans

Activation of canonical Wnt signaling is sufficient and necessary to inhibit the migration of the QR.pa neuroblast of C. elegans. To identify targets of canonical Wnt signaling involved in this process we isolated QR lineage descendants by FACS from control (mab-5(gk670) mutant) animals and animals expressing a constitutively active, N-terminally truncated form of BAR-1/β-catenin in the QR lineage using the egl-17 promoter (ΔN-BAR-1Q). FACS isolation was based on the expression of mCherry and GFP markers under the control of wrt-2 and egl-17 promoters, respectively. We performed differential gene expression analysis to identified genes that are differentially expressed between control and constitutive activation of canonical Wnt signaling. mRNA profiles of QR descendants from control (mab-5(gk670) mutant) and ΔN-BAR-1Q larvae were generated from pools of 2000 cells with CEL-seq protocol, in triplicate, using Illumina HiSeq 2500.