Fig 2: a7-nAChR levels show less variability along the basal forebrain axis following different treatments. (A,C,E) Representative total protein staining used as loading control. (B,D,F) Representative immunoblots and bar graphs indicating the a7-nAChR levels along the three BF subdivisions. (A) Total protein membrane subsequently immunoblotted for a7-nAChR. (B) The receptor did not show any rostro-caudal difference between the two conditions [F(5, 55) = 0.3227, p = 0.8972, two-way ANOVA]. (C) Total protein membrane subsequently immunoblotted for a7-nAChR. (D) No interaction was observed between conditions and anatomical subdivisions [F(5, 35) = 1.503, p = 0.2139, two-way ANOVA]. (E) Total protein membrane subsequently immunoblotted for a7-nAChR (F) No change was detected along the BF anatomical plane after T15 administration [F(5, 35) = 0.2600, p = 0.9318, two-way ANOVA]. Protein changes were indicated as mean ± SEM. n = (hemisections per condition, rats) in (A,B) are (36, 12); in (C,D) (24, 8); in (E,F) (24, 8).

Fig 3: AChE-peptide and a7-nAChR levels and their interaction showed an age-dependent pattern during postnatal brain development. (a,c) Representative immunoblots showing the protein expression in six hemislices from one randomly-chosen animal per age. (b,d) Bar graphs indicating the protein profile related to four different ages. (a,b) AChE-peptide expression was not significantly affected across ages; however, its levels revealed a decreased tendency from early postnatal to young adult animals (trend analysis post-hoc test). (c,d) a7-nAChR levels were strongly decreased in an age-related manner (trend analysis post-hoc test). In particular, a significant difference was detected when comparing P7 rats to the other groups (Bonferroni’s post-hoc test). (e) Representative Co-IP blot showing the molecular interaction between T14 and the nicotinic receptor. (f) The complex T14/a7-nAChR showed a significant decreasing trend across ages (trend analysis post-hoc test) and also a clear change between the two extreme ages (Bonferroni’s post-hoc test). All values are normalised against the P7 group, considered as the control. Asterisks indicate a difference between P7 and the other groups (Bonferroni’s post-hoc test), while hashes represent the linear trend among groups (trend analysis post-hoc test). Data are indicated as the mean ± SEM. n = (hemislices, rats) is (24, 4). *,# p < 0.05, ****,#### p < 0.0001.