Fig. 7. Single-cell RNA-seq analysis of the nanos2^−/− mutant reveals the absence of primordial germ cells and gametes.

(A) Single-cell transcriptomic map of an individual nanos2^−/− mutant juvenile polyp merged with WT adult tissue libraries. (B) Library contribution to the map in (A) illustrates a similar contribution of the mutant to all principal cell populations with the exception of germ cells and uncharacterized cell type S3 (black outline). (C) Distribution of mutant cells (red) across the entire dataset (gray), and the cluster of putative stem cells (pSCs) and germ cells identifies the absence of germ cells and differentiated gametes in nanos2^−/− mutants. (D) Expression dot plot of pSCs, germ cells, and neuroglandular derivative cell populations. Detection of gene expression in at least 5% of the cell population is illustrated. Germ cell marker expression (nanos2, piwi1, and vasa2) shows all three genes are coexpressed and elevated within the WT germ cell populations, as well as detected in both mutant and WT pSCs. nanos2 is also broadly detected across all populations in the WT samples and is partially coexpressed with markers of early neuroglandular progenitors nanos1, soxC, and soxB(2), whose expression is unaffected in the mutant.