Fig 1: Pannexin 1 functions upstream of epithelial Nras and Bcas2 to drive injury-induced proliferation in the regenerating epithelium.(a) Schema of experimental protocol for RNAseq analysis of purified lung epithelium from Panx1+/+ and Panx1-/- mice after naphthalene epithelial injury with (b) heat map of differentially regulated genes, units representing log2 scaled counts. (c) CRISPR–Cas9 genetic deletion of Panx1, Nras and Bcas2 was performed in BEAS-2B epithelial cells (confirmation at transcript and protein level) with (d) proliferation of these cell lines measured by EdU incorporation over 24h, displayed relative to control (n=5, assessed by Kruskal-Wallis test with Dunn's multiple comparisons test). (e) Measurement of Nras (n=7) and Bcas2 (n-5) in BEAS-2B cells after overnight treatment with recombinant human amphiregulin (rAreg) (assessed by Wilcoxon signed-rank test). (f,g) Generation of Nras and Bcas2 zebrafish morphants (or control sequence morphants) for assessment of regeneration rate at 48h post-tailfin transection (n=12-18 separate animals, two separate experiments, assessed by t-test) and (h) tailfin proliferation analysed by EdU incorporation (n=14-39 per group, assessed by t-test). *p<0.05, **p<0.01.