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. 2022 May 18;25(6):104426. doi: 10.1016/j.isci.2022.104426

Figure 3.

Figure 3

Severely impaired migration of CCR10+ lymphocytes into the skin of total CCL27-knockout mice

(A) Diagram of CRISPR targeting in total CCL27-knockout mice.

(B) Genomic PCR identifying wild-type (+/+), heterozygous (+/−), and homozygous (−/−) total CCL27-KO mice. Band sizes of wild-type and knockout CCL27 alleles are 484 and 607bp, respectively.

(C) Immunofluorescent α-CCL27 antibody staining images on 10μm skin sections from CCL27+/+ and CCL27−/− mice. Staining with a nonspecific goat IgG antibody or the secondary antibody (2′ Ab) only is included as controls. The dashed lines mark the surface of the skin. N = 3 mice for each genotype.

(D) FC analysis of CD45+ cells from the skin and spleen of CCL27+/+ and CCL27−/− recipient mice two days after receiving transfer of sLN cells of CCR10+/EGFP mice. Donor-derived CCR10(EGFP)+ lymphocytes are gated on CD45.1+ cells. N = 6 mice for each group.

(E) Total numbers (#) of donor cells and percentages (%) of donor cells that express CCR10(EGFP) in the skin and spleen of CCL27+/+ and CCL27−/− recipient mice. N = 6 mice for each group. ∗p < 0.05; ∗∗p < 0.01; ns: no significant difference. Paired one-tailed t-test.