Fig. 4: Combined effect of E-cadherin loss-of-function and pro-inflammatory activation supports a 2-hit model for E-cadherin loss in the neural crest. | Nature Communications

Fig. 4: Combined effect of E-cadherin loss-of-function and pro-inflammatory activation supports a 2-hit model for E-cadherin loss in the neural crest.

From: Neural crest E-cadherin loss drives cleft lip/palate by epigenetic modulation via pro-inflammatory gene–environment interaction

Fig. 4

a twist ISH in control, LPS + ATP low (250ug/ml LPS + 0.1 mM ATP), E-cadherin morpholino low (ECAD MO low; 2 ng ECAD MO) and LPS + ATP low + ECAD MO low treated samples. Neural crest migration length is evidenced by black brackets. b Quantification of neural crest migration lengths in control (n = 10), LPS + ATP low (n = 10), ECAD MO low (n = 10) and LPS + ATP low + ECAD MO low (n = 10), showing significant reduction of neural crest migration length in the combined LPS + ATP low + ECAD MO low conditions (p = 0.002, Two-way ANOVA). c cdh1 mRNA levels showing significant reduction in LPS + ATP low + ECAD MO low conditions when compared to controls (n = 4 per group; p = 0.008. Two-way ANOVA). d Scratch assays using hiNCCs in wild type, TNFa exposure, heterozygous E-cadherin knockout (ECAD KO) and combined ECAD KO + TNFa conditions evidencing scratch areas at 0 h (yellow line) and 24 h (blue line) after scratch in each condition. e Quantification of scratch percentage of closure in wild type, TNFa exposure, ECAD KO and ECAD KO + TNFa’in hiNCCS after 24 h (n = 9 per group). TNFa significantly reduces scratch closure in comparison to wild type (p < 0.0001) and combining ECAD KO + TNFa significantly reduces scratch closure in comparison to ECAD KO (p = 0.0305) and to TNFa (p = 0.0008). Two-way ANOVA. f qPCR quantification of CDH1 expression in wild type, TNFa, ECAD KO and ECAD KO + TNFa conditions of hiNCCs (n = 7 per group). TNFa significantly reduces CDH1 expression in comparison to wild type (p < 0.0001) and combining ECAD KO + TNFa significantly reduces CDH1 expression in comparison to ECAD KO (p = 0.0236) and to TNFa (p < 0.0001). Two-way ANOVA. Boxplots centre is the median, with bounds representing the 25th and 75th percentile, with and whiskers as minima to maxima. Source data are provided as a Source Data file. *p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001.

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