Fig. 5

5-Nonyloxytryptamine (5-NL) induces differential gene expression and activates the AMPK pathway in tumor cells. B16.GP33 cells were treated with 5-NL (3 µM) for 18 h and RNA was assessed using RNA-seq analysis. A A Volcano Plot of the fold change gene distribution is shown. B-C GSEA analysis of pathways altered in 5-NL treated B16.GP33 cells is shown with arrow pointing up indicating pathway activation and arrow pointing down indicating downregulation. Fold changes in individual genes in select pathways are shown in C (n = 4). (D) Level of AMPK phosphorylation (Thr172) was assessed using immunoblot analysis in B16 and MC38 cells treated with 3 µM of 5-NL at the indicated time points quantified in the right panel (black frames indicate cropped immunoblot; n = 7–9). Error bars indicate SEM; *P < 0.05 as determined by a one-way ANOVA with a Dunnett’s post-hoc test