Figure 6. TSLP secreted by mutant^GFP epidermal cells induces hyper‐proliferation of non‐mutant^Tom keratinocytes.

• A
  Experimental design for induction of mutant^GFP KCs in vitro and neutralization of secreted TSLP by anti‐TSLP. Primary keratinocytes were cultured from the skin of Junb^lox/lox; c‐Jun^lox/lox; Rosa^Ki/+; and K5cre‐ERT^+/+ mice. 50% of mutant^GFP KCs were induced by infection with Cre Adenovirus, the non‐infected cells were non‐mutant^Tom KCs. Adeno‐empty was used as control. EdU was added for 5 h to label proliferating Tomato⁺ cells in 6‐day co‐culture. Quantification was performed by confocal imaging analysis.
• B
  TSLP production in conditioned media quantified by ELISA three days after infection with Ad‐empty or Ad‐cre. n = 3 independent experiments. Data represent mean ± SD. Statistical significance ***P < 0.001 (Student's two‐tailed t‐test relative to control group). See Appendix Table S2 for exact P‐values.
• C
  Percentage of EdU⁺ Tomato⁺ keratinocytes after blocking with anti‐TSLP. Anti‐TSLP neutralizes the hyper‐proliferation of non‐mutant^Tom KCs in vitro after infection with Ad‐cre. n = 3 independent experiments, six fields per experiment. Data represent mean ± SD. Statistical significance *P < 0.05 (two‐way ANOVA and Bonferroni post‐test).
• D
  Gene expression of Tslpr after TSLP neutralization is reduced. n = 3 independent experiments. Data represent mean ± SD. Statistical significance *P < 0.05 (Student's two‐tailed t‐test relative to control group). See Appendix Table S2 for exact P‐value.
• E
  Gene expression of IL‐7ra after TSLP neutralization is increased. n = 3 independent experiments. Data represent mean ± SD. Statistical significance *P < 0.05 (Student's two‐tailed t‐test relative to control group). See Appendix Table S2 for exact P‐value.
• F
  Gene expression of VEGFα after TSLP neutralization is reduced. n = 3 independent experiments. Data represent mean ± SD. Statistical significance *P < 0.05, **P < 0.01 (Student's two‐tailed t‐test relative to control groups). See Appendix Table S2 for exact P‐value.
• G
  Experimental design for TSLP neutralization in vivo. TSLP neutralization was performed by triple intradermal injections of 20 μg of anti‐TSLP or IgG to the right ears of DKO* and DKO*¹⁵ mice 5 days after psoriasis‐like induction. EdU was added into mice by IP injection 2 h before euthanasia.
• H
  Representative images of treated ears after 9 days of anti‐TSLP or IgG treatment.
• I, J
  Longitudinal analysis of ear thickness (I) and trans‐epidermal water loss (TEWL) (J) in DKO* mice at different time points during psoriasis‐like disease progression. Data represent mean ± SD. n = 4. Statistical significance *P < 0.05 (two‐way ANOVA and Bonferroni post‐test).
• K
  Percentage of EdU⁺ epidermal cells (CD45⁻CD49f⁺) after blocking with anti‐TSLP in DKO* and DKO*¹⁵ mice. DKO* n = 4 and DKO*¹⁵ n = 3 per group. Data represent mean ± SD. Statistical significance *P < 0.05 (two‐way ANOVA and Bonferroni post‐test).
• L
  Representative immunofluorescence images of ear sections from control or DKO* mice treated with IgG or anti‐TSLP and stained for Ki67. Ki67 expression was reduced in mice treated with anti‐TSLP (n = 3).

Source data are available online for this figure.