Skip to main content
. 2023 Nov 13;16:1241420. doi: 10.3389/fnmol.2023.1241420

Figure 15.

Figure 15

L-NAME treatment does not affect p75NTR levels in aged BFCNs. Aged rat BFCNs (DIV18-21) were treated with 1 mM  L-NAME for 24 h prior to analysis of p75NTR levels using immunocytochemistry. p75NTR is shown in green and DAPI is shown in blue. (A,E) DIV8 BFCNs treated with vehicle; (B,F) DIV21 BFCNs treated with vehicle; (C,G) DIV21 BFCNs treated with L-NAME; (D) Quantification of the images shown in A–C. No differences in p75NTR levels were observed between any of the groups at the cell bodies. For both vehicle-treated groups, n = 30 images. For L-NAME, n = 29 images. For each group, images were obtained from three chambers in three independent experiments. One-way ANOVA, p > 0.05. The highest two points of the DIV18-21 vehicle and the highest point of the DIV7-9 vehicle were detected as outliers using ROUT (Q = 1%). Removal of these outliers did not change the significance of the comparison (p = 0.061). (H) Quantification of the images shown in E–G. Aged BFCNs had greater p75NTR levels at the axon terminals compared to young BFCNs regardless of treatment condition. For all groups, n = 30 images obtained from three chambers in three independent experiments. The lowest point of the DIV7-9 vehicle was detected as an outlier using ROUT (Q = 1%). Removal of this outlier did not affect the significance of the comparison between the vehicle-treated groups (p = 0.01) but changed the comparison between DIV7-8 vehicle and DIV18-21  L-NAME from p = 0.002 to p = 0.0006. One-way ANOVA and post hoc Tukey test, *p < 0.05, **p < 0.01. Error bars: SEM. DIV, days in vitro. (I,J) Immunocytochemistry results were confirmed using western blotting for p75NTR in DIV21 BFCNs treated with either vehicle or 1 mM  L-NAME. (I) Representative western blot comparing relative intensity of p75NTR expression normalized to β-actin expression. 16.8  μg of total protein was loaded in each BFCN sample well; 33.6  μg total protein from p75NTR exon III knockout and wildtype mouse hippocampal tissues were loaded as negative and positive controls, respectively. (J) There were no differences in p75NTR expression between BFCNs treated with L-NAME (n = 4) and vehicle treatment (n = 5; p = 0.95). Error bars: SEM. DIV, days in vitro. Scale bars: 10 μm.