Fig 1: Transcriptomic Signature of In Vitro Differentiating Primary Human Airway Epithelial Cells. (A) Time course profile of expression of select genes from clusters plotted in Figure 1B . Left panel: microarray gene expression; right panel: hemi-nested qPCR validation of microarray data for days 0, 2, 8 and 28 post-ALI (same hSAEpC donor [donor #1]). (B) qPCR plots illustrating comparable patterns in expression of OMG, KRT14, STC1, BPIFA1, PLA2G7, TXNIP and S100A7 in three additional healthy human airway epithelial cell (hAEpC) donors, consistent with (A). Top left (donor #2) and right (donor #3) panels: human bronchial epithelial cells [hBEpCs]: bottom left panel: hSAEpCs (donor #4). (C) Heatmap illustration of change in expression of genes in (A) and (B) across all tested donors (donors #1–4). OMG, oligodendrocyte myelin glycoprotein; KRT14, keratin 14; STC1, stanniocalcin 1; BPIFA1, BPI fold containing family A member 1; PLA2G7, phospholipase A2 group VII; TXNIP, thioredoxin interacting protein; S100A7, S100 calcium binding protein A7.
Fig 2: BPIFA1 Is a Secretory Biomarker of Differentiating Human Airway Epithelial Cells. Confirmation of BPIFA1 expression at protein level (secreted basolaterally into culture media) from six hAEpC donors (bronchial epithelial cells derived from two different donors [designated as yellow (donor #5) and red (donor #6)], and small airway epithelial cells derived from four separate donors [designated as purple (donor #7), orange (donor #8), blue (donor #9), and green (donor #10)]) with n = 4-7 replicates per donor. Data were analyzed by non-parametric Mann-Whitney test. ***p < 0.001, ****p < 0.0001, n.s., not significant.
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