Fig. 2: Ligand endocytosis mediates temperature-dependent Notch activity. | Nature Communications

Fig. 2: Ligand endocytosis mediates temperature-dependent Notch activity.

From: Temperature sensitivity of Notch signaling underlies species-specific developmental plasticity and robustness in amniote brains

Fig. 2

a NICD-dependent Notch reporter activity in chick neural progenitors cultured at 30 °C or 37 °C (mean + SD, n = 3 biologically independent samples in each group). b Relative expression levels of genes encoding Notch signaling components in chick neural progenitors cultured at 30 °C or 37 °C (mean + SD, n = 3 biologically independent samples in each group). c Overexpression of Notch1 (NotchWT) or mutant Notch (NotchL468A) receptors in developing chick neural progenitors. d Notch reporter activities in control, NotchWT, and NotchL468A overexpressing samples cultured at 30 °C or 37 °C (mean + SD, n = 3 biologically independent samples in each group). e Schematic drawing of Dll1/4 and Dll1/4ΔC; the latter lacks the intracellular domain (ICD) required for ubiquitylation and endocytosis. f, g Notch reporter activity in controls and Dll1ΔC- (f) or Dll4ΔC (g) -overexpressing neural progenitors cultured at 30 °C or 37 °C [mean + SD, n = 3 (f) and 4 (g) biologically independent samples in each group]. h Notch reporter activity in control and dynasore-treated neural progenitors cultured at 30 °C or 37 °C (mean + SD, n = 3 biologically independent samples in each group). i Notch reporter activity in control and dominant-negative dynamin 2 (DN-dynamin 2) overexpressing neural progenitors cultured at 30 °C or 37 °C (mean + SD, n = 3 biologically independent samples in each group). j Notch reporter activity in control and Dll4-overexpressing neural progenitors cultured at 37 °C (mean + SD, n = 4 biologically independent samples in each group). k Schematic illustration of GFP or Dll4 overexpression in the developing chick pallium. I Distribution of Sox2 and/or GFP-positive cells in the developing chick pallium in control and Dll4 electroporated samples. m The proportion of Sox2-negative and GFP-positive cells in controls and Dll4-overexpressing chick pallium at 37 °C (means + SD, n = 3 biologically independent samples in each group). Two-sided, unpaired t-test for a, j, and m (p-values are indicated in each graph), two-sided, negative bimodal distribution for b (p-values were adjusted for multiple comparisons by using Benjamin–Hochberg method, FDR of Dll1 is not true), ordinary one-way ANOVA for d and f–i (p-values were adjusted by Tukey test for multiple comparisons). Scale bar: 25 µm.

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