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. 2023 Sep 8;12(18):2231. doi: 10.3390/cells12182231

Figure 4.

Figure 4

Regulation of NICD and SNW1 binding via HDAC6-mediated deacetylation. HPMEC-Im transduced with lentivirus containing sc shRNA or HDAC6 shRNA were treated with 50 ng/mL VEGF for 45 min. Immunoprecipitation (IP) with NICD antibody was performed with subsequent Western blot analysis carried out with an acetylated antibody and HDAC6 antibody on immunoprecipitated protein to quantify changes in NICD acetylation and HDAC6-NICD binding (A). Densitometric quantification of Western blot images in Figure 4A is shown graphically (B), n = 5 independent experiments; * p < 0.05. Similarly, NICD acetylation and NICD-SNW1 binding were assessed after IP with NICD antibody (C), with the results presented graphically (D), n ≥ 4 independent experiments, * p < 0.05. SNW1 acetylation and NICD-SNW1 binding were measured through immunoblotting after IP with SNW1 antibody (E), and densitometry analysis shown graphically (F), n = 5 independent experiments and * p < 0.05. (G) SNW1 binding to RBPJ binding sites on the promoters of HES1 and HEY2 was quantified using Chromatin IP (ChIP) with SNW1 antibody. n = 4 independent experiments, * p < 0.05.