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. Author manuscript; available in PMC: 2019 May 31.
Published in final edited form as: Cell. 2018 May 31;173(6):1356–1369.e22. doi: 10.1016/j.cell.2018.03.051

Figure 6. NOTCH2NL paratypes interact with NOTCH receptors and enhance Notch signaling.

Figure 6

(A) Co-immunoprecipitation of NOTCH2 and NOTCH2NL analyzed by immunoblot. IP=Immunoprecipitation, N2NL=NOTCH2NL (B,C) Co-transfection of NOTCH2-GAL4 and NOTCH2NL in the pGL3-UAS reporter assay in co-culture with U2OS cells (B) or U2OS-JAG2 cells (C), n = 24 in 4 experiments. (D) Immunoblot showing immunoprecipitated NOTCH2NL in the medium of mouse ESCs ectopically expressing NOTCH2NL. (E) Effect of NOTCH2NL-conditioned medium in NOTCH reporter assay, n = 7 in 2 experiments. One-way anova with Tukey’s HSD (* p<10−4). (F–G) NOTCH reporter assay using the NOTCH1 receptor in conjunction with NOTCH2NL. (H–K) Assessment of common NOTCH2NL variants in reporter assay. (H–I) n = 12 in 2 experiments. (J–K) n = 6 in 1 experiment, t-test with Holm-Bonferroni correction (* p<0.05, ** p<10−3), error bars indicate SD. Data is analyzed using Two-way anova with Tukey’s HSD (* p<10−4, ** p<10−8, *** p<10−12). Error bars indicate SEM, unless otherwise specified. See also Figure S6.