Fig. 2.

The transcript levels of cytokines are increased and tissue-specific induction of cytokines is achieved within the induced ins-CETI-PIC3 model system. (A) Schematic for the experimental procedure. All treatments were performed at 72 h post-fertilization (hpf). For B, C and D, RNA was subsequently isolated from the whole bodies of at least 15 embryos in each clutch and all analysis was performed on whole-body samples. (B) Dose response to 0, 0.5, 1 and 2.5 µg/ml doxycycline is shown for tnfa, ifng1 and il1b, and H2B-GFP. (C) Time response to 5 µg/ml doxycycline is shown for up to 12 h after treatment with doxycycline. All experiments for D and E were performed in ins-CETI-PIC3 experimental embryos and clutch-mate control Tg(ins:cre) or CETI-PIC3 embryos treated with 5 µg/ml doxycycline for 48 h. (D) The levels of tnfa, ifng1 and il1b, and H2B-GFP are all increased after doxycycline induction in the ins-CETI-PIC3 embryos compared to Tg(ins:cre) and CETI-PIC3 control embryos. n=4. *P<0.05 (one-way ANOVA). (E) After crossing the CETI-PIC3 line to the Tg(ins:cre) line, β cell-specific induction of the cytokines occurred, as seen by the H2B-GFP signal and Tnfa staining only in the Insulin-positive cells (arrows).