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. 2021 Feb 24;14(2):dmm046300. doi: 10.1242/dmm.046300

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© 2021. Published by The Company of Biologists Ltd

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.

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Fig. 6. — NOTCH3^L1519P hyperactivates expression of Notch downstream genes. (A) Western blot analysis of PDGFRB expression in HMFΔN2 cells expressing wild-type NOTCH3 (WT) or NOTCH3^L1519P (L1519P) compared to the parental HMFΔN2 cells (Ctrl). The graph shows PDGFRB expression normalized to β-actin using ImageJ. (B) Quantitative real-time PCR analysis of the Notch downstream target genes NOTCH3, HES1, HEY1, NRARP and PDGFRB from wild-type or NOTCH3^L1519P-expressing HMFΔN2 cells. (C) Western blot analysis of PDGFRB, AKT and p42/44 MAPK phosphorylation upon PDGF-BB stimulation as indicated. The blot with p42/44 MAPK was stripped and re-probed with the β-actin antibody for a loading control. Relative levels were normalized to β-actin using ImageJ (n=3). Statistical significant was analyzed from three experiments using paired Student's t-test, *P<0.1, **P<0.01, ***P<0.001; ns, not significant.