Journal: Cell Biology and Toxicology

Article Title: N-nitrosodimethylamine induces cytochrome P450 activation but not epithelial remodeling in well-differentiated human airway epithelial cells

doi: 10.1007/s10565-026-10181-4

Figure Lengend Snippet: NDMA exposure does not elicit significant remodeling of airway epithelial cells. In well-differentiated HBE cells exposed to NDMA (up to 5 mM) apically for 4 h daily, we detected airway epithelial differentiation markers by RT-qPCR ( A - C ) and IF staining ( D - E ) at 1, 3, and 5 days post-exposure. A-C: NDMA exposure showed no significant difference in mRNA expression of (A) TP63, (B) MUC5AC, and (C) FOXJ1 compared with control. Each symbol represents an independent donor (mean ± SD, n = 3 HBE cell donors). *p < 0.05 versus vehicle control (0 mM). Two-way ANOVA with Dunnet’s multiple comparison test. D-E: Representative IF images demonstrate that NDMA did not cause changes in the number of differentiated cell types, as determined by IF staining of P63, MUC5AC for goblet cells (D), and FOXJ1 (E). Scale bars = 100 μm

Article Snippet: Cells were permeabilized with 0.2% Triton X-100 in DPBS for 10 min and blocked with 1% BSA and 10% normal goat serum for 1 h. The HBE cells were stained for F-actin (1:40, Alexa fluor 488-Phalloidin, ThermoFisher Scientific) for 30 min or for proteins of, as follows: E-cadherin (1:400, 3195S, Cell Signaling Technology), ZO-1 (1:100, 33–9100, Thermo-Fisher Scientific), MUC5AC (1:100, MA5:12,178, ThermoFisher Scientific), FOXJ1 (1:100, 14–9965-82, ThermoFisher Scientific), P63 (1:500, 13,109, Cell Signaling Technologies).

Techniques: Quantitative RT-PCR, Staining, Expressing, Control, Comparison