Fig 2: MLKL-mediated necroptosis triggers skin inflammation in OTULINE-KO mice.a Kaplan–Meier plot depicting survival of mice of the indicated genotypes. b Graph depicting macroscopic skin score of mice of the indicated genotypes. The dots in the graphs represent individual mice. Mean ± s.e.m. is shown for each group of mice in all graphs. Statistical significance was determined using Kruskal–Wallis test (one-sided). c Representative images of skin sections from 3-week-old mice of the indicated genotypes, stained with H&E (n = 4) or immunostained with anti-CC3 (n = 3), anti-CC8 (n = 3) or anti-Ki-67 (n = 3) antibodies. Scale bars: H&E = 100 µm; CC3, CC8, Ki-67 = 50 µm. d Representative images from skin sections from 3-week-old mice of the indicated genotypes, immunostained with anti-K10, anti-K14 (n = 4), anti-K6 (n = 3), anti-F4/80 (n = 3) and anti-Gr-1 (n = 3) antibodies and counterstained with DAPI (DNA stain). Scale bars: K10, K14, K6 = 20 µm; F4/80, Gr-1 = 30 µm. e Graphs depicting relative mRNA expression of the indicated genes in RNA from whole-skin tissue of 3-week-old mice of the indicated genotypes, measured by qRT-PCR. The dots in the graphs represent individual mice. Mean ± s.e.m. is shown for each group of mice in all graphs. Statistical significance was determined using Kruskal–Wallis test (one-sided). Control mice include littermates that do not express K14-Cre. Source data for a, b, e are provided as a Source data file.

Fig 3: Mice lacking OTULIN in keratinocytes develop skin inflammation.a Photographs of lesional skin from OTULINE-KO mice at the age of 3 and 14 weeks and representative images from skin sections from 3- or 14-week-old mice from OTULINE-KO, stained with haematoxylin and eosin (H&E). Images shown are representative of OTULINE-KO mice (n = 25). Scale bars: H&E = 200 µm. b Representative images of skin sections from 3-week-old mice of the indicated genotypes, stained with H&E (n = 10) or immunostained with antibodies against cleaved caspase 3 (CC3) (n = 6), cleaved caspase 8 (CC8) (n = 6) or Ki67 (n = 6). The epidermis in the sections stained for Ki-67 is indicated by a dotted line, arrows point to hair follicles. Scale bar H&E = 100 µm; CC3, CC8, Ki-67 = 50 µm. c Representative images of skin sections from 3-week-old mice of the indicated genotypes, immunostained with antibodies against keratin 10 (K10), keratin 14 (K14) (n = 6), keratin 6 (K6) (n = 6), F4/80 (n = 6) and Gr-1 (n = 6) and counterstained with DAPI (DNA stain). Scale bars: K10, K14, K6 = 20 µm; F4/80, Gr-1 = 30 µm. d Heatmap of gene expression data of OTULINE-KO lesional or non-lesional skin tissue compared to skin from littermate OtulinFL/FL control mice. Columns represent gene expression profiles from 3-week-old mice (n = 5), showing genes with an absolute log2(FoldChange) of =1, and an uncorrected p value of =0.05. Uncorrected p values are used on purpose to include a maximal number of genes in the plot while excluding clearly irrelevant genes. The statistical test producing the p values is Wald test for the absence of differential expression, i.e. for a log2(FoldChange) of zero. It is computed by function nbinomWaldTest of the Bioconductor R package DESeq2, based on a negative binomial general linear model of the gene counts. Control mice include littermates that do not express K14-Cre. Source data for d are provided as a Source data file.

Fig 4: MyD88-dependent signalling contributes to the development of skin pathology in OTULINE-KO mice.a Kaplan–Meier plot depicting survival of mice of the indicated genotypes. b Graph depicting macroscopic skin score of mice of the indicated genotypes. The dots in the graphs represent individual mice. Mean ± s.e.m. is shown for each group of mice in all graphs. Statistical significance was determined using Kruskal–Wallis test (one-sided). c Representative images from skin sections from 3-week-old mice of the indicated genotypes, stained with H&E (n = 5) or immunostained with anti-CC3 (n = 5), anti-CC8 (n = 5) or anti-Ki-67 (n = 5) antibodies. Scale bars: H&E = 100 µm; CC3, CC8, Ki-67 = 50 µm. d Representative images from skin sections from 3-week-old mice of the indicated genotypes, immunostained with anti-K10, anti-K14 (n = 5), anti-K6 (n = 5), anti-F4/80 (n = 5) and anti-Gr-1 (n = 5) antibodies and counterstained with DAPI (DNA stain). Scale bars: K10, K14, K6 = 20 µm; F4/80, Gr-1 = 30 µm. e Graphs depicting relative mRNA expression of the indicated genes in RNA from whole-skin tissue of 3-week-old mice of the indicated genotypes, measured by qRT-PCR. The dots in the graphs represent individual mice. Mean ± s.e.m. is shown for each group of mice in all graphs. Statistical significance was determined using Kruskal–Wallis test (one-sided). Control mice include littermates that do not express K14-Cre. Source data for a, b, e are provided as a Source data file.

Fig 5: TNFR1 signalling in keratinocytes causes skin inflammation in OTULINE-KO mice.a Kaplan–Meier plot depicting survival of mice of the indicated genotypes. b Graph depicting macroscopic skin score of mice of the indicated genotypes. The dots in the graphs represent individual mice. Mean ± s.e.m. is shown for each group of mice in all graphs. Statistical significance was determined using Kruskal–Wallis test (one-sided). c Representative images of skin sections from 3-week-old mice of the indicated genotypes, stained with H&E (n = 6) or immunostained with anti-CC3 (n = 5), anti-CC8 (n = 5) or anti-Ki-67 (n = 5) antibodies. Scale bars: H&E = 100 µm; CC3, CC8, Ki-67 = 50 µm. d Representative images of skin sections from 3-week-old mice of the indicated genotypes, immunostained with anti-K10, anti-K14 (n = 5), anti-K6 (n = 5), anti-F4/80 (n = 3) and anti-Gr-1 (n = 3) antibodies and counterstained with DAPI (DNA stain). Scale bars: K10, K14, K6 = 20 µm; F4/80, Gr-1 = 30 µm. e Graphs depicting relative mRNA expression of the indicated genes in RNA from whole-skin tissue of 3-week-old mice of the indicated genotypes, measured by qRT-PCR. The dots in the graphs represent individual mice. Mean ± s.e.m. is shown for each group of mice in all graphs. Statistical significance was determined using Kruskal–Wallis test (one-sided). Control mice include littermates that do not express K14-Cre. Source data for a, b, e are provided as a Source data file.