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. 2022 May 17;23(6):e53791. doi: 10.15252/embr.202153791

Figure EV2. Construction and identification of keratinocytes IL‐38‐specific knockout mice and effect of keratinocyte‐specific IL‐38 deficiency on epidermal barrier.

Figure EV2

  1. Construction of Il‐38‐loxP (Il‐38f / f ) mice.
  2. Typical genetic cross scheme of keratinocyte IL‐38‐specific knockout (K14Cre /+ ‐Il‐38f / f ) mice.
  3. PCR analysis of WT mice (at 317 and 245 bp) and Il‐38‐loxP mice (at 387 and 373 bp).
  4. PCR analysis of keratin 14 promoter directing expression of Cre recombinase(left) and deletion of IL‐38 (right).
  5. Relative expression of IL‐38 in the epidermal splits of Il‐38f / f (n = 4) and K14Cre /+ ‐Il‐38f / f (n = 4) mice.
  6. Representative western blot bands indicating IL‐38 in the epidermal splits of Il‐38f / f and K14Cre /+ ‐Il‐38f / f mice.
  7. Appearance of Il‐38f / f and K14Cre /+ ‐Il‐38f / f mice.
  8. Cumulative body weight changes in Il‐38f / f (n = 12) and K14Cre /+ ‐Il‐38f / f (n = 12) mice in 60 days.
  9. Representative histological photographs of skin stained with hematoxylin‐eosin (H&E) from Il‐38f / f (n = 5) and K14Cre /+ ‐Il‐38f / f (n = 5) mice. Scale bars represent 100 μm. The graph shows the quantification of epidermal thickness.
  10. Skin barrier‐dependent dye exclusion assay using toluidine blue in Il‐38f / f mice (n = 5) and K14Cre /+ ‐Il‐38f / f littermate (n = 5) at birth.
  11. TEWL assay measured on ventral surface of newborn Il‐38f / f mice (n = 6) and K14Cre /+ ‐Il‐38f / f littermate (n = 6).
  12. Representative micrographs of cornified cell envelopes from Il‐38f / f (n = 4) and K14Cre /+ ‐Il‐38f / f (n = 4) mice. Scale bars represent 100 μm. The graph shows the number of cornified cell envelopes per square millimeter.
  13. Representative immunohistochemical staining micrographs of CD45 in the skin of Il‐38f / f (n = 5) and K14Cre /+ ‐Il‐38f / f (n = 5) mice. Scale bars represent 100 μm. The graph shows average intensities of CD45 per high‐powered field.

Data information: Error bars represent the mean ± SD. All data are biological replicates. ns, not significant; *P < 0.05; **P < 0.01; ***P < 0.001; P values were calculated using Student’s t‐test.