Journal: Cell Communication and Signaling : CCS

Article Title: Reduced SUMOylation impairs NOTCH3 signaling and cell survival in the pathogenesis of CADASIL

doi: 10.1186/s12964-025-02556-7

Figure Lengend Snippet: Impairment of cell survival and proliferation in NOTCH3 mutant cell lines and primary cultured cerebral VSMCs. A and B Representative images depicting cell death ( A ) and proliferation ( B ) in HEK293 cells with transient overexpression of empty vector (EV), NOTCH3-WT (N3-WT) and the NOTCH3 mutant (NOTCH3-R90C, N3-R90C). C and D Representative images of cell death ( C ) and proliferation ( D ) in HEK293 cells stably overexpressing empty vector (EV), NOTCH3-WT (N3-WT) and mutant NOTCH3 (NOTCH3-R90C and NOTCH3-R544C, N3-R90C and N3-R544C). E and F Representative images of cell death ( E ) and proliferation ( F ) in primary cultured cerebral VSMCs from WT and NOTCH3-R545C (HE and HO) mice. Cell survival is assessed by Live & Dead staining. Green indicates the live cells and red indicates the dead cells. Cell proliferation is identified by Ki67 (red) immunocytochemistry. DAPI (blue) indicates the nuclear location. G - L Statistical data on the cell survival and proliferative rate in HEK293 cells with transient ( G and H ) and stable ( I and J ) overexpression of the NOTCH3 mutant, as well as in primary cultured cerebral VSMCs ( K and L ). Experiments were independently repeated six times. One-way ANOVA with Tukey post hoc test. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: Thereafter, the VSMCs were incubated with a rabbit anti-Ki67 primary antibody (1:300, Proteintech, 27309-1-AP) overnight at 4°C.

Techniques: Mutagenesis, Cell Culture, Over Expression, Plasmid Preparation, Stable Transfection, Staining, Immunocytochemistry