Extended Data Fig. 4: The recovery of endogenous full-length Cdk3 by correcting the Cdk3 pre-mature mutation.
From: CDK3 induces neuronal death and brain atrophy in Alzheimer’s disease
(a) Generation of a Cdk3-ps (Term-Try187) point mutation mouse model was achieved using CRISPR/Cas9 technology. (b) The relative mRNA levels of Cdk3-ps were analyzed in the brains of Cdk3-ps(A561G/A561G) mouse (Abbreviated as Cdk3(561DM)) and littermate controls using two primer pairs, namely Cdk3-ps-PP1 and Cdk3-ps-PP2, followed by RT-PCR analysis. (c) Western blotting with a c-terminal antibody against Cdk3 revealed the presence of full-length Cdk3 protein in cortical lysates from mutated mice (Cdk3(561DM)), which was absent from their littermate controls. (d-e) Neuronal morphology was detected after treatment with Aβ in primary neuron cells from WT and Cdk3(561DM) mice. e represents statistical analysis of total branch length. Scale bar: 50 μm. (f) After treatment with Aβ, synaptic markers (Snap25, Synaptophysin and Psd95) were assessed in primary neurons from WT and Cdk3(561DM) mice. (Sample number: n = 8 cases for WT and Cdk3(561DM) respectively in b, n = 4 cases for WT and Cdk3(561DM) respectively in c. n = 21-33 neurons per groups, 107 neurons in all in e. Data represent mean ± SEM. p < 0.05 indicates significance between the two indicated groups. b. e: two-way ANOVA with Tukey post hoc test.).
