Fig. 2
From: Structural determinants for activation of the Tau kinase CDK5 by the serotonin receptor 5-HT7R
Expression and functional characterization of 5-HT7R mutants with impaired Gs coupling. A Scheme of receptor mutants. Substitution mutants E325G, K327S and E325G;K327S possess single or double exchanged amino acids in the ICL3, whereas ΔR395 lacks the carboxy-terminal domain. B, C Expression of HA-tagged 5-HT7R WT or mutants in N1E-115 cells. Representative Western blot (B) and quantification (C). The 5-HT7R levels were normalized to GAPDH and are shown as mean ± SD (N = 6, one-way ANOVA, Dunnett’s multiple comparisons, *** p < 0.01). D Representative confocal images showing expression of eYFP-tagged 5-HT7R WT or mutants. Corresponding intensity profiles are shown below. Scale bar: 10 μm. See also Additional file 6. E N1E-115 cells were transfected with cAMP fluorescence resonance energy transfer-based biosensor CEPAC and indicated constructs of eYFP-tagged 5-HT7R. Basal cAMP levels are shown. Signals are normalized to cells transfected with 5-HT7R WT (3 ≤ N ≤ 7, one-way ANOVA, Dunnett’s multiple comparison, * p < 0.05). F Representative traces showing cAMP response at the single cell level after stimulation with 10 µM of serotonin (5-HT). G Graphs show cAMP response amplitude relative to pretreatment (3 ≤ N ≤ 7, one-way ANOVA, Dunnett’s multiple comparisons, *** p < 0.001)