Fig. 4: Loss of REST delays pulmonary neuroendocrine cell fate transition in response to naphthalene injury.

a Single-cell RT-qPCR analysis of Rest expression in pulmonary neuroendocrine cells (PNECs) and non-NE lung epithelial cells from normal lungs of two Chga-GFP adult mice (n = 87 cells). b Timeline of the PNEC lineage tracing and naphthalene-induced lung injury experiment. Tom, Tomato fluorescent reporter. c Whole-mount fluorescence images of left lung lobes 3 weeks after injury. Images representative of n = 2 independent experiments. Scale bar, 5 mm. d, e Quantification (d) of normal neuroepithelial bodies (NEBs) and PNEC outgrowths after injury as in (c) with each column representing a single mouse left lung lobe and each data point a tomato-positive cluster in that mouse. A Wilcoxon rank sum test, two-sided, was conducted on the combined PNEC outgrowths from all the mice in each respective group that are above the threshold size of the non-injured controls (n = 107 for Rest^+/+, n = 146 for Rest^fl/fl). Median area (e) of all Tom⁺ clusters per mouse as in (d) (n = 2 mice for corn oil controls, 4 mice for naphthalene-treated Ascl1^CreER/+;Rest^+/+ and 6 mice for naphthalene-treated Ascl1^CreER/+;Rest^fl/fl). Unpaired t-test, data represented as mean ± s.d. f, g Percentage (f) and absolute number counted (g) of Tom⁺CC10⁺ clusters over total number of Tom⁺ clusters at various timepoints after naphthalene injury (n = number of clusters as indicated in table (g) totaled from representative left lung lobe sections of 3–4 mice per timepoint). Fisher’s exact test, two-tailed. See also Supplementary Fig. 7. Source data are provided as a Source Data file.