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. Author manuscript; available in PMC: 2020 Mar 18.
Published in final edited form as: Curr Biol. 2019 Feb 28;29(6):1064–1072.e8. doi: 10.1016/j.cub.2019.01.076

Figure 3. daf-18 has a diffuse cellular focus within the somatic gonad.

Figure 3.

All dauers contain daf-7(e1372); daf-18(ok480), markers, excisable daf-18(+), and Cre drivers.

(A) Strategy to generate excision mosaics. The conditional allele arTi179, daf-18 [loxP>daf-18>loxP], restores normal dauer development in a daf-18(0); daf-7 background. Tissue-specific expression of Cre recombinase using the single-copy insertion drivers shown (box) generates excision mosaics in which daf-18(+) is removed from the DTCs [hlh-12p], all somatic gonad cells [ckb-3p], or variably in all/most or no somatic gonad cells [hlh-1p] because of transient expression in MS granddaughters [52, 53]. None of the Cre drivers used caused excision in the germline. For more details of excision patterns observed with these drivers, see STAR Methods. daf-18(0) excision mosaics were obtained using a cross strategy (box) that (i) eliminated potential maternal contribution of daf-18(+) by providing the DAF-18-expressing loxP>daf-18(+)>loxP floxed locus through the male and (ii) required a single excision event involving the daf-18 [loxP>daf-18>loxP] locus present in F1 heterozygous progeny, so that all GFP-positive DTCs lacked daf-18(+) activity. For further details about generation of transgenes and additional observations about the full effects of the drivers used see STAR Methods.

(B) SGB and GSC progression in excision mosaics. The excision pattern induced by each driver shown was confirmed by both GFP expression from the excision event involving the single daf-18 [loxP>daf-18>loxP] locus (see above) in the mosaics scored and analysis of heIs105, an established recombination reporter [25] (STAR Methods). SGB, somatic gonad blast cell; DTC, distal tip cell; GSC, germline stem cell. SGB division progression was inferred by substantially increased number of cells expressing ckb-3p::mCherry::H2B and concomitant expansion by DIC, and germline divisions were scored by DIC (see Figure 1 B,C). (***) P ≤ 0.001 by two-tailed Fisher’s exact test.

See also Figure S3.