Fig 1: EREG rescues the growth defects in Sod1 deficient organoids.A Western blot analysis of EREG and SOD1 expression using the separated stromal compartment from Sod1f/f and Sod1f/f; Vil-creERT2 mice after 24 h post 5 consecutive tamoxifen injections. n = 4 mice per genotype. B Western blot analysis of SOD1 expression in Sod1f/f and Sod1f/f; Vil-creERT2 organoids after 5 days of tamoxifen with or without 0.5 μg ml-1 of recombinant EGF/AREG/EREG induction. n = 3 mice per genotype. C Representative images of organoids after 5 days of tamoxifen with or without EGF/AREG/EREG induction. n = 6 wells per group; n = 3 mice per genotype. Red arrowhead indicates dead organoids. D Percentage of organoids with 0, 1, 2, 3, or ≥4 crypts formed after 5 days of tamoxifen with or without EGF/AREG/EREG induction. Data represent mean ± SEM; n = 6 wells per group; n = 3 mice per genotyping. Statistical significances were tested by two-way ANOVA. *P < 0.05, ***P < 0.001. Number of organoids counted: n = 249 (Sod1f/f), n = 287 (Sod1f/f; Vil-creERT2), n = 307 (Sod1f/f; Vil-creERT2 + EGF), n = 309 (Sod1f/f; Vil-creERT2 + AREG), n = 272 (Sod1f/f; Vil-creERT2 + EREG) organoids per group; one of three experiments. E Percentage of dead organoids assayed after 5 days of tamoxifen with or without EGF/AREG/EREG induction. Data represent mean ± SEM; n = 6 wells per group; n = 3 mice per genotyping. Statistical significance was tested by one-way ANOVA. ***P < 0.001. Number of organoids counted: n = 393 (Sod1f/f), n = 475 (Sod1f/f; Vil-creERT2), n = 537 (Sod1f/f; Vil-creERT2 + EGF), n = 532 (Sod1f/f; Vil-creERT2 + AREG), n = 413 (Sod1f/f; Vil-creERT2 + EREG) organoids per group; one of three experiments. All results are representative of at least three independent experiments.
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