Figure 5
From: Identification of Novel Inhibitors of DLK Palmitoylation and Signaling by High Content Screening
Ketoconazole significantly decreases DLK-mediated phospho-cJun activation in primary neurons. (A) Dorsal Root Ganglion (DRG) neurons were treated at 7 Days in vitro (DIV 7) with 2.5 µM Ketoconazole overnight or 20 µM 2BP for 2 h prior to a 2.5 h NGF withdrawal in presence of the indicated compound. Cells were lysed in SDS-PAGE loading buffer and levels of endogenous DLK, phospho-cJun (p-cJun) and tubulin were detected by western blot as indicated. The secondary antibody used on the p-cJun blot also weakly recognizes residual anti-NGF IgG used as part of the trophic factor deprivation (indicated by asterisk). (B) Quantification of phospho-cJun normalised to –NGF vehicle-treated cells. Two-way ANOVA indicates significant effects of interaction (p = 0.0071), NGF (p = 0.0026) and Ketoconazole (p = 0.0001). The effect of Ketoconazole in DRGs undergoing NGF withdrawal was also significant as determined by the Bonferroni post-test (p < 0.01). Error bars represent SEM.
