Figure 3. Engineered mDASCs possessed normal stem cell properties and enhanced antimicrobial potency.

• A–C
  Detection of LL‐37 expression in the engineered mDASCs by immunofluorescence (A), real‐time quantitative PCR (B), and Western blot (C). Scale bar, 50 μm. BF, bright field. n = 10. Error bars, SEM.
• D
  Anti‐Krt5 (red) and anti‐P63 (green) immunostaining of WT‐ and LL‐37‐mDASC colonies. Scale bar, 70 μm.
• E
  Stem cell colony‐forming efficiency of WT‐ and LL‐37‐mDASCs during five serial passages. n = 6. Error bars, SD.
• F
  Representative 3D organoid culture of mDASCs with expression of type I alveolar cell markers (Aqp5 and Pdpn). Left panels, bright‐field imaging of 3D organoids. Right panels, immunofluorescence of organoid sections. Scale bar, 20 μm.
• G
  Co‐culture of bacteria with DASCs shows antimicrobial effects in dose‐dependent manner. Initial additions of PAO1 were 0.1 × , 0.5 × and 1 × 10⁴ CFU, respectively. Co‐culture duration, 6 h. n = 4. Error bars, SEM. MOI, multiplicity of infection.
• H
  Co‐culture of bacteria with DASCs shows antimicrobial effects in time‐dependent manner. Initial concentration of PAO1 was 1 × 10⁴ CFU. MOI = 1. n = 3. Error bars, SEM.
• I, J
  Preincubation of cells with anti‐LL‐37 antibody, but not IgG control, significantly reduced anti‐PAO1 (I) and anti‐Escherichia coli (J) effects of LL‐37‐mDASCs. Initial dose of bacteria was 10³ CFU. Co‐culture duration, 18 h. n = 4 in (I) and n = 3 in (J). Error bars, SEM.

Data information: Statistics for graphs: unpaired two‐tailed t‐test (B), two‐way ANOVA followed by Sidak's test (G, H) and one‐way ANOVA followed by Tukey's test (I, J). *P < 0.05; **P < 0.01; ****P < 0.0001.Source data are available online for this figure.