Fig. 5: Deletion of SOX9 in human beta cells.

A The differentiation strategy of human embryonic stem cells to pancreatic beta cells, with DOX addition. B Immunostaining for SOX9 at day 11 of differentiation in the absence (left panel) or presence (right panel) of DOX. The experiment was carried out at least three independent times. C Strategy for deleting SOX9 in endocrine progenitors. D eBCs generated from the iSOX9KO line in the absence (left panels) or presence (right panels) of DOX. GFP fluorescence is shown in green in the upper panel and bright field images are shown in the lower panels. The experiment was carried out at least three independent times. E Flow analysis on iSOX9KO cells differentiating either in the presence or absence of DOX. n = 3 for the NO DOX samples and n = 2 for the DOX samples were used for costaining C-PEP with either NKX6.1, NEUROD1 or CHGA, and for the GCG measurements. For the total C-PEP measurements, n = 6 for the NO DOX samples and n = 4 for the DOX samples were used. F Dynamic Insulin secretion in control eBCs (No DOX, grey tracing, n = 6) and SOX9 knockout eBCs (DOX, orange tracing, n = 6). G Area under the curve calculated for the first phase of insulin secretion as shown in (F). H Total insulin content measured in control (grey bar, No DOX, n = 4 independent experiments) or knockout (orange bar, DOX, n = 3 independent experiments) eBCs. Error bars represent S.E.M. using a two-tailed Student’s t test, *p < 0.05, **p < 0.005. Source data are provided as a Source Data file.