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. 2019 Apr 16;9(6):1933–1943. doi: 10.1534/g3.119.400162

Figure 1.

Figure 1

sfGFP approach in C. elegans germline A) Design of genetic loci. GFP1-10 was inserted into a locus that is permissive for germline expression on chromosome II under the control of a germline specific promoter and 3′UTR. GFP11 fragments were inserted via CRISPR to tag proteins at their endogenous location. B) Schematic representation of sfGFP approach for expression of germline proteins. C) RT-PCR on RNA extracted from gfp1-10 worms indicates that gfp1-10 is expressed. Actin PCR serves as a positive control to confirm RNA extraction was successful (expected RT-PCR size = 151bp). GFP1-10 RT-PCR product is expected to be 170bp, while standard PCR of genomic DNA produces a 270bp sized product. The expected product size for GFP1-10 cDNA is only observed in the GFP1-10 sample. D) Expression of GFP1-10 alone does not lead to GFP fluorescence, similar to wild-type (N2) worms. E) Quantification of fluorescence of GFP1-10 strain shows that there is no fluorescence in the absence of GFP11 tag. Nuclear intensity calculated as Intensity/Area(µm2), for pre-meiotic and mid-pachytene regions. Scale bars 2μm.