Fig. 6

Fig. 6

Insulin knockdown enhances regeneration of β cells. (A) Schematic of β cell ablation and regeneration experiments. Control and insaMO-injected insa:CFP-NTR embryos were treated with MTZ from 52 to 80 hpf, recovered in fresh egg water for 1 day, and analyzed at ~100 hpf (1 day post ablation; dpa). (B–C′) Merged (B,C) and single channel (B′,C′) confocal projections of regenerated 100 hpf control (B,B′) and insaMO-injected (C,C′) islets immunostained for Gcg (red), CFP (β cells, green), and DNA (blue). Arrows indicate the expansion of strongly double positive Ins+ Gcg+ cells. (D–E′) Merged (D,E) and single channel (D′,E′) confocal projections of 100 hpf control (D,D′) and insaMO-injected (E,E′) islets immunostained for CFP (green) and DNA (blue). All islets expressed zygotically-injected H2B-RFP (red) to mark DBCs. Arrows indicate the expansion of double positive H2B-RFP+ Ins+ cells; this indicates conversion of non-β endocrine cells into β cells. (F) Quantification of insa+, Gcg+, and insa+ Gcg+ cells in 1 dpa control (n=17) and insaMO-injected (n=22) islets. (G) Quantification of H2B-RFP+ insa+ and H2B-RFP^-insa+ regenerating β cells in 1 dpa control (n=17) and insaMO-injected (n=13) islets. Student′s t-test was used in G to determine statistical significance.