Figure 6: Changes in the transcript levels of procollagens during the epithelial recanalization, measurement of the elastic modulus.

(A-C) Transcript levels of Procol1A1, Procol1A2 and Procol3A1 in control and drained vocal folds 24h, 48h and 72h PS. For qPCR analysis, control embryos collected at E16.5 (24h PS) were used as controls for ΔΔCT data normalization. Data are presented as the average of two biological replicates (n=6) ± standard error of the mean, as three mouse larynges were pulled together for one biological replicate to maximize RNA yield. A Student’s T-test, ANOVA and post hoc Tukey test were performed to calculate significance in the gene expression levels between control and drained embryos at each time point and between time points. A p-value < 0.05 (*) was considered significant. (D) Measurements of the elastic modulus in control and drained VF. 24h and 48h PS in drained samples the lamina propia was significantly softer than in control VF. However, at 72h PS the stiffness of the lamina propria in drained samples significantly increased, which correlates with the increased expression of collagens in the lamina propria. A Welch ANOVA and a post hoc Games-Howell test were used to calculate significance, as they do not make assumptions about equal variance between groups. A p-value < 0.05 (*) was considered significant