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Fig. 5
TBK1/IKK? inhibitors accelerate restoration of ?-cell function by selectively increasing the number of ?-cells. (A?C) Confocal images of [Tg(ins:CFP-NTR)s892; Tg(ins:Kaede)jh6] larvae at 48 hpa, concurrently treated with EdU and DMSO (A), amlexanox (B), or PIAA (C), respectively, from 0?48 hpa, stained for Somatostatin (blue). The number of Somatostatin-expressing ?-cells that incorporated EdU (purple arrows) did not increase in TBK1/IKK?-I-treated recovering larvae (B, C) compared to DMSO-treated larvae (A). (D) The percentage (mean?±?SD) of ?-cells that incorporated EdU at 48 hpa (in A-C; 12.9?±?2.0% (DMSO), 12.4?±?4.7% (amlexanox), and 15.1?±?4.3% (PIAA)). Cells in 20 planes of confocal images from 10 individual larvae were counted per condition. (E?G) Confocal images of [Tg(ins:CFP-NTR)s892; Tg(ins:Kaede)jh6] larvae at 48 hpa, concurrently treated with EdU and DMSO (E), amlexanox (F), or PIAA (G), respectively, from 0?48 hpa, stained for Glucagon (blue). The number of Glucagon-expressing ?-cells that incorporated EdU (blue arrows) did not increase in TBK1/IKK?-I-treated recovering larvae (F, G) compared to DMSO-treated larvae (E). (H) The percentage (mean?±?SD) of ?-cells that incorporated EdU at 48 hpa (in E-G; 6.2?±?2.4% (DMSO), 8.0?±?1.9% (amlexanox), and 10.9?±?3.4% (PIAA)). Cells in 20 planes of confocal images from 10 individual larvae were counted per condition. (I) Free-glucose levels (mean?±?SD) during ?-cell regeneration in non-ablated wild type, DMSO-treated recovering, and TBK1/IKK?-I-treated recovering larvae. At 7 dpf (equivalent to 72 hpa), free-glucose levels were significantly lower in PIAA-treated recovering larvae (blue line, 457.7?±?28.8 pmol/larva) than in DMSO-treated larvae (purple line, 719.3?±?42.2 pmol/larva). *P?<?0.05. n?=?30 larvae (3 pools of 10 larvae) per data point.