| |
Figure 6. Insm1 and Rfx6 are necessary for ectopic beta cell promotion by Ngn3-GR. In situ hybridizations for insulin expression in all panels. Ngn3-GR mRNA (1,800 pg) was coinjected with a control mismatch morpholino (mMO) or morpholinos targeting insm1 or rfx6 mRNA, activated with Dex for 4 h and fixed at Stage 32. (a,b) Insm1 mMO (20 ng) had no effect on endogenous insulin expression in either ex (n = 8/8) or +Dex (n = 12/17) treated embryos. (c,d) Injection of Insm1 MOs (2 morpholinos, 20 ng each) abolished endogenous insulin expression in ex embryos (n = 7/7) as well as ectopic insulin expression in +Dex embryos (n = 15/15). (e,f) Rfx6 mismatch morpholino (25 ng) did not affect endogenous or ectopic insulin expression in ex (n = 9/9) or +Dex embryos (n = 19/41). (g,h) Antisense Rfx6 morpholino (25 ng) inhibited endogenous insulin expression in −Dex embryos (n = 33/33) as well as ectopic insulin expression in +Dex embryos (n = 46/46). |
