Fig. 2.

Leukocytes from the gene-edited fetus express CD18 without the signal peptide. (A) Leukocytes from the gene-edited fetus contain the Q(‒5)G substitution in both alleles of CD18. Sequencing data shows the presence of glycine-encoding GGA codon in the CD18 DNA from the gene-edited fetus (a1) and glutamine-encoding CAG codon in the CD18 DNA from the wild-type CD18 DNA (a2). (B) CD18 expression on leukocytes from the gene-edited fetus is comparable to that on leukocytes from a wild-type calf. CD18 expression on the leukocytes from the gene-edited fetus (b1) or a wild-type calf (b2) was determined by flow cytometry. The blue and red histograms represent binding of anti-CD18 antibodies and isotype-matched control antibodies, respectively. The black histogram represents unstained cells. (C) CD18 expressed on leukocytes from the gene-edited fetus lacks the signal peptide. The presence of signal peptide on CD18 expressed on the leukocytes from the gene-edited fetus (c1) or a wild-type calf (c2) was determined by flow cytometry. The magenta and blue histograms represent binding of chicken anti-signal peptide serum and unimmunized chicken serum, respectively. The red histogram represents unstained cells. Leukocytes from the wild-type calf expressed CD18 containing the signal peptide, whereas the leukocytes from the gene-edited fetus expressed CD18 without the signal peptide. Results of one representative experiment of two are shown.