Fig 2. Confirmation by qPCR of the effect of NIST and vitamin D on expression of cytokine genes in HBEC cultures.

(A) Gene mRNA expression at 4 hours relative to 18S and the unstimulated Control condition in HBEC cultures stimulated with 50μg/ml NIST in the presence/absence of 100nM 1,25(OH)₂D3. IL6, n = 7; IL8, n = 7; IL24, n = 6; CFS2, n = 8; CXCL10, n = 6; TGFB2, n = 6. (B) mRNA expression at 24 hours. IL6, n = 9; IL8, n = 7; IL24, n = 6; CFS2, n = 8; CXCL10, n = 6 (one outlying replicate excluded); TGFB2, n = 6. All repeated-measures ANOVAs and post-tests with Bonferroni corrections as shown. VC; vehicle control for NIST. Statistical significance as follows: *, p ≤ 0.05; **, p ≤ 0.01; ***, p ≤ 0.001; ****, p ≤ 0.0001.