Fig. 5: Modelling SOX9-mediated salivary gland development in organoid cultures.
a, A subset of putative clones and a pooled population from each condition were analysed using junction PCR and confirmed on-target integration of the cassette into the AAVS1 locus. b, The clones from each condition were tested containing a SOX2-specific gRNA. Samples were cultured in the presence or absence of Dox (2 μM) for 4 d and analysed using real-time RT-PCR. The data are normalized to ribosomal protein L27 and are presented as means ± s.d. (n = 3 biological replicates). Statistical significance was determined by one-way ANOVA followed by Tukey’s post-hoc test. c, The clones from each condition were tested containing SOX9-specific gRNAs. Samples were cultured in the presence or absence of Dox (2 μM) from differentiation day 80 and analysed for dCas9 (left) and SOX9 (right) expressions using RT-qPCR. The data are normalized to ribosomal protein L27 and are presented as means ± s.d. (n = 3 biological replicates). Statistical significance was determined by one-way ANOVA followed by Tukey’s post-hoc test. d, Salivary gland-like branching structures observed in the aggregate. Arrowheads indicate salivary gland-like structures under the control conditions (top and middle). Salivary gland-like structures were not detected under Dox-dependent SOX9 knockdown conditions (bottom). Scale bars, 500 μm. e, Efficiency of salivary gland-like structure formation under each condition (n = 24 organoids (per treatment); two independent experiments).
