Supplementary Figure 2: EGFR deletion in EGFR^ΔMx∗ mice and cytokine production on DEN injection.

(a) ELISA showing IL-1β in the supernatant of primary hepatocyte cultures 4 h after incubation with increasing amounts of DEN in vitro. (Primary hepatocyte isolates of EGFR^f/f (n = 3) and EGFR^Δhep (n = 3)). (b) Release of IL-1β and IL-1α to the supernatant of cultured primary hepatocytes of EGFR^f/f, EGFR^Δhep, and EGFR^ΔMx mice after incubation with TNFα quantified by ELISA. n.d. = not detectable. Result of two pooled independent experiments is shown. For each experiment hepatocytes isolated from 2 livers per genotype were pooled and analysed as 4 technical replicates (primary hepatocytes of n = 4 mice were analysed in total for each genotype). (c) Western Blot analysis of EGFR in livers of EGFR^f/f and EGFR^ΔMx∗ mice. (d) Representative livers of EGFR^f/f and EGFR^ΔMx∗ mice 46 weeks after tumour initiation. Scale bar indicates 1 cm. (e) Tumour mass (left, EGFR^f/f (n = 10) and EGFR^ΔMx∗ (n = 9)), area (middle, EGFR^f/f (n = 7) and EGFR^ΔMx∗ (n = 9)), and number (right, EGFR^f/f (n = 7) and EGFR^ΔMx∗ (n = 9)). Two pooled independent experiments. Data (a,b) represent mean ± s.d. Data (e) represent mean ± s.e.m. Student’s t-test for independent samples and unequal variances was used to assess statistical significance (^∗P < 0.05, ^∗∗P < 0.01, ^∗∗∗P < 0.001). Original data are provided in Supplementary Table 1.