Figure 6.

Impaired Airway Regeneration after Ablating Lgr6⁺ Cells

(A) Schematics of the Lgr6-EGFP-CreERT2;R26-Tom lineage-tracing experiment after airway injury.

(B) Representative confocal images showing extent of club cell injury or repair with (PBS, top) or without Lgr6⁺ cells (DT, bottom) after naphthalene administration by IF analysis: Tdtomato (for Lgr6, red); Ki67 (white); CCSP (cyan); and DAPI (blue). Arrow, Ki67⁺ airway epithelial cells; arrowhead, Ki67⁺Lgr6⁺ cells.

(C–E) Quantification of naphthalene injury repair and expansion of Lgr6⁺ cells in lungs with or without Lgr6⁺ cells. (C) For club cell analysis, the percentage of DAPI-positive bronchiolar cells also positive for CCSP is assessed at indicated time points. (D and E) The mean percentage of club cells or lineage-labeled Lgr6⁺ cells that co-express Ki67 is assessed at indicated time points.

(F) Experimental scheme of depleting Lgr6-expressing cells in co-culture of Scgb1a1⁺ cells.

(G and H) Representative phase-contrast images (G) and colony forming efficiency (H) of primary lung organoids in (F).

(I) qPCR analysis for the expression of Fgf10 in isolated lineage-labeled Lgr6⁺ cells from naphthalene-treated lungs. Shown is normalized to Gapdh.

(J and K) Colony forming efficiency (J) and quantification of each distinct colony type (K) of lung organoids with addition of DT and Fgf10 at day 7 in cultures. Data presented are the mean of experiments from three individual mice per group (D, E, and I) or the mean of three independent experiments with triplicate wells (H, J, and K).

Error bars indicate SD (^∗p < 0.01; ^∗∗p < 0.005). The scale bars represent 50 μm (B) and 500 μm (G). See also Figure S5.