Fig. 4
Mirodenafil inhibits transcriptional activity and homodimerization of GR. A TF profiling plate array was performed after treatment with Aβ and mirodenafil in HT-22 cells. B GR Dual-Luciferase reporter gene assay after treatment with Aβ and mirodenafil in HT-22 cells (n ≥ 5). C GR Dual-Luciferase reporter gene assay after co-treatment with dexamethasone and mirodenafil or CORT-108297 in HEK293 cells. D Western blot analysis for GR after co-treatment with Aβ42 and mirodenafil in HT-22 cells. E PolarScreen™ Glucocorticoid Receptor (GR) Competitor Assay to determine the IC50 values of dexamethasone and mirodenafil that directly bind to the full length of recombinant GR protein. F Co-immunoprecipitation of Myc-tagged GR with GFP-tagged GR in HT-22 cells after treatment with dexamethasone, mirodenafil (Mir), sildenafil (Sil), tadalafil (Tad), and vardenafil (Vad). G Quantifications of co-immunoprecipitation of GR homodimers after treatment with dexamethasone and PDE5 inhibitors using ImageJ software. Data are presented as the mean ± SEM; *P < 0.05, **P < 0.01, ****P < 0.0001. All statistical comparisons were performed relative to Aβ42- or dexamethasone-treated cells. Statistical significance was assessed by one-way ANOVA followed by Bonferroni’s post hoc test for multiple comparisons