Fig 2. Electroporation of zygotes incompatible with pronuclear injection.

(A) Average ratio of harvested zygotes with polar bodies from natural mating with visible pronuclei (+) and without visible pronuclei (-) from 47 independent experiments. Total number of embryos analyzed was 6368 and 3314 for zygotes with and without visible pronuclei, respectively. (B) Representative RFLP analysis for evaluation of HDR efficiency in Nphs2-targeted blastocysts upon EEZy of zygotes without visible pronuclei. PCR controls from untreated blastocysts (WT) and without DNA template (H₂O) and RFLP controls from untreated blastocysts (WT) are depicted. (C) Quantification of the genotype from three independent experiments using zygotes without visible pronuclei are compared to EEZy of all zygotes from the experiments depicted in Fig 1B and 1D, right columns. (D) Percentage of developed blastocysts from zygotes after EEZy from (C). Box plot represents median, boxes equal 25 to 75 percentiles, whiskers include all values. Remaining data are means ± standard deviation. ***p < 0.001, ns = non-significant. N = total number of embryos analyzed.