Supplementary Figure 2: Effect of Sorafenib on oesophageal epithelium.

(a–c) Proliferation following Sorafenib treatment. (a) Protocol. Animals were treated with Sorafenib or vehicle only (Control) for 10 days and injected with EdU (purple arrow) 1 h before being culled. (b,c) Box plots showing dose response to Sorafenib of percentage of EdU + (b) and Ki67 + (c) basal cells in epithelial wholemounts. Values are % positive cells/field in random fields of view from duplicate animals at each time point. Number of fields sampled in b was 0 mg and 1 mg, 12 fields, 10 mg, 9 fields, 50 mg 11 fields, 100 mg 10 fields, and in c 0 mg, 10 fields, 1 mg and 10 mg, 9 fields, 50 mg 8 fields, 100 mg 10 fields. Box indicates 25th and 75th centiles, centre line is median, whiskers indicate range. (d–h) Characterization of epithelium after 28 days Sorafenib treatment. (d) Protocol. (e) Confocal image of epithelial cryosection stained for Cryosection stained for P-AKT (Ser473, green) and basal marker ITGA6 (white) and DAPI (blue). Scale bar, 50 μm. Arrow, cells positive for P-AKT. Image representative of sections from 3 animals/group. (f) Mean percentage of basal cells staining positive for P-AKT (^∗P = 0.011 by two tailed t-test, n = 4 animals/group). (g) Typical confocal images of epithelial cryosections stained for basal keratinocyte marker KRT14 (green), suprabasal marker KRT4 (magenta), basal marker ITGA6 (white) and DAPI (blue). Scale bar, 50 μm. Arrow, cells double positive for KRT4 and KRT14 (inset). (h) Mean percentage of cells staining positive for activated Caspase 3, n = 3 animals per group. See Supplementary Table 4 for source data for b,c and f.