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. Author manuscript; available in PMC: 2023 Apr 25.
Published in final edited form as: Dev Cell. 2022 Apr 13;57(8):1068–1080.e6. doi: 10.1016/j.devcel.2022.03.013

Figure 3.

Figure 3.

Establishing safety and efficiency of the dTAG system in post-implantation stages.

(A) (Left) Schematic of testing dTAGv-1 safety in post-implantation development in vivo. Wild type pregnant females were given IP injection of 35mg kg−1 dTAGv-1 at 6.5, 8.5, 10.5 days post conception (dpc). (Right) Resulting healthy pups at term were counted.

(B) Quantification of HA mean signal intensity in E6.5 embryos 4hrs after heterozygous NelfbdTAG/+ pregnant females received dTAGv-1 or dTAG-13 injection. Each dot represents an embryo. HA signal was normalized to Hoechst, then vehicle injected litters.

(C) Immunofluorescence images of E6.5 embryos retrieved 4hrs after pregnant females received IP injection of dTAGv-1 or dTAG-13. Nuclei are labeled by Hoechst. Scale bars, 50μm.

(D) Quantification of HA mean signal intensity in E7.5 embryos 1,2,3, or 4hrs after heterozygous NelfbdTAG/+ pregnant females received dTAGv-1 injection. Each dot represents an embryo. HA signal was normalized to Hoechst, then vehicle injected litters.

(E) Immunofluorescence images of E7.5 embryos retrieved 1,2,3 and 4hrs after pregnant females received IP injection of dTAGv-1. Nuclei are labeled by Hoechst. Scale bars, 50μm.

(F) Quantification of HA mean signal intensity in E9.5 embryos 4 and 8hrs after heterozygous NelfbdTAG/+ pregnant females received one dTAGv-1 injection, or two separated by 2hrs and followed for 6hrs after the second for a total 8hrs from first injection. Each dot represents an embryo. HA signal was normalized to Hoechst, then vehicle injected litters.

(G) Immunofluorescence images of E9.5 embryos retrieved 8hrs after pregnant females received two IP injections of dTAGv-1. Nuclei are labeled by Hoechst. Scale bars, 500μm.

For all experiments, maximum intensity projection (MIP) is shown in images. Plots show each data point with group mean and interquartile range. Student t-test was used to determine significance. Statistical significance is classified based on p-value as: n.s. > 0.05, * < 0.05, ** < 0.01, *** < 0.001, **** < 0.0001.