Figure 3. Normal Defense and Mucus Secretory Cells Are Reprogrammed by IL-13 into a Distinct Pathologic Mucus Secretory State.

(A) Immunofluorescence (IF) labeling of ALI-differentiated human AEC cultures in top-down (top) and side (bottom) views show control cultures (left) dominated by SCGB1A1⁺ (green) and MUC5AC⁻ (red) cells transformed into cultures (right) with prominent co-staining (yellow) after acute IL-13. n = 3 HTEC donors (T73, T76, T79); scale bar, 30 mm; DAPI nuclei labeling (blue).

(B) Heatmap shows shared functions between pairs of control and IL-13-dominant secretory populations (c5/c6, c7/c8, and c9-control/c9-IL-13). Mean scaled expression of population-defining DEGs within enriched functionally annotated sets is shown.

(C) Scatterplot compares responses to IL-13 in defense (y) and mucus (x) secretory cells. Points, expression log fold changes with IL-13; filled, differentially expressed in both populations; unfilled, differentially expressed in one population; colored, belonging to specified function categories.

(D) TFs whose expression significantly changes across a pseudotime trajectory transitioning from baseline to IL-13-stimulated cell states. Left: TFs increasing (top) or decreasing (bottom) across pseudotime. Right: scaled normalized expression (points) and smoothed trends (lines) for exemplar TFs across pseudotime-ordered cells. Point color, population as in (B); line color, defense-secretory (black), mucus-secretory (red).

(E) Dot plots show shifts in level (dot color) and ubiquity (dot size) of expression for genes "activated" (top) or "suppressed" (bottom) by IL-13 in one or both secretory populations. Functional groups as in (C). Gene label font color: gold, defense; brown, mucus (population with strongest effect is shown). Same direction responses in both population pairs are indicated by asterisk (significant) or point (not significant).

See also Figure S2.