Fig. 3: Expression and activity of MMPs during hANO development.

A Heatmap shows expression levels of MMP7, MMP9, MMP10 and MMP13 based on RNA-seq data. B mRNA levels of MMP7, MMP9, MMP10, and MMP13 were analyzed in hANOs at D10, D17 and D24 by qPCR assay; the experiment was independently performed in five organoid lines from five different donors. C Protein levels of MMP7, MMP9, MMP10, and MMP13 were analyzed in hANOs at D10, D17 and D24 by Luminex assay; the experiment was independently performed in four organoid lines from four different donors. D Pan-MMP activity was analyzed in hANOs at D10, D17, and D24, Pan-MMP activity was analyzed in hANOs at D10, D17, and D24; NC-1 and NC-2 were negative controls in which the organoids treated with culture medium and assay buffer, respectively; while PC was the positive control in which the organoids treated with APMA pre-treated MMP9 recombinant protein; RFU relative fluorescence units; data present the mean with SEM, n = 5 experiments per condition; the experiment was independently performed in five organoid lines from five different donors. The Friedman test was used in comparison analysis in (B) and (C); the 2way ANOVA with Tukey’s multiple comparison test was performed in (D).