Fig. 6: A novel acquired mutation in the 16th EGF domain of NOTCH3 contributes to metastatic phenotype of HNSCC cells.
From: Identification of acquired Notch3 dependency in metastatic Head and Neck Cancer
We utilized CRISPR/Cas9 methodology and performed genome editing of UT-SCC-74B line, correcting the point mutation identified in Notch3 gene. CRISPR corrected cell line is labeled as 74B* in all figures. a An increase in confluency in parental lines and CRISPR edited clone was accessed over time using Incucyte Zoom Instrument. The growth rate of 74B* line differs from UT-SCC-74B (p value < 0.0001) and resembles the growth rate of UT-SCC-74A. b–d The expression of Notch ligands, target genes and markers of EMT in CRISPR edited cell line was accessed by qPCR comparing to parental UT-SCC-74A and UT-SCC-74B cells. e Cells were plated in 96-well plates and transfected with siRNA against Notch3. Confluency of cells before and 5 days after transfection was accessed using Incucyte Zoom Instrument; the increase in confluency was calculated and normalized to control (mock transfected) for each cell line. siRNA for EG5 was used as a positive control for transfection. All the data in this figure is presented as mean ± SEM.
