Single-cells transcriptomes of mouse developing forelimbs

In order to try and evaluate Hoxd transcript heterogeneity during limb development, we produced single-limb cell transcriptomes of different origins, to see whether the apparently homogenous behavior in Hox gene transcriptional program as observed upon large-scale analyses was confirmed at the cellular level. We report here thatHoxd genes transcripts are present in various combinations in different limb cells and discuss the importance of these results in our understanding of how Hoxd genes are regulated and how their global functions are achieved in these structures. Single-cells transcriptomes from Hoxd11::GFP forelimbs with micro-dissection of autopod and zeugopod at embryonic day 12 (E12.5). Dissociated single-cells were obtained from eight Hoxd11::GFP forelimbs micro-dissected at E12.5. Cells with the highest level of GFP fluorescence (top 20%) were sorted using an Astrios cell sorter with a 100-μ m nozzle. 75bp reads were uniquely mapped to the latest Mus Musculus reference genome (mm10) and the ERCC sequences using bowtie2 (REF: doi:10.1038/nmeth.1923) in local mode.