Nicotine treatment lowered CYP2A protein levels in the lower and upper lung in AGM study 1
Most AGM lung samples, loaded at 75 μg, were within the linear range of the CYP2A, β–actin and Coomassie blue signals from the standard curve of pooled control AGM lung (described in the Methods) on each immunoblot (Figure 5A and 5B). However, a few samples had CYP2A detection which were below the linear detection level and were thus reassessed after loading at 125 μg. A representative blot from AGM study 1 is shown in Figure 5A and 5B; representative AGM lung membranes from each group (n=3) were loaded at 75 μg.
The Nicotine group, vs the Control group, had significantly lower levels of CYP2A protein in the lower (4.03-fold) and upper (3.74-fold) lung (Figure 5C and 5D). Blot-to-blot adjusted (as shown in Figure 5C–D) and unadjusted data provided virtually identical significant fold group differences (Supplementary Table 1).
CYP2A protein levels in the lower and upper lung were highly correlated (r2=0.88), within individual AGMs (Figure 5E). There were no significant differences in CYP2A/β–actin levels between the lower and upper lung of the Control group or Nicotine group.