Figure 3—figure supplement 2.

Figure 3—figure supplement 2.

(A) Cartoon showing the rationale of using mo^SPtcf7l2 that targets intron 4/exon five splice site boundary blocking the splicing machinery and making it skip to the next splicing acceptor site in exon 6. (B) RT-PCR (top panel) and western blot (WB, middle and bottom panels) performed using RNA and proteins extracted from 24hpf zebrafish embryos injected with: control morpholino (first lane), mo^SPtcf7l2 (moSP, second lane), and mo^ATGtcf7l2 (moATG, third lane). cDNA was amplified using set ‘a’ primers (Figure 1A, 5’F1 vs Splice R1, Materials and methods). Anti human Tcf7l2 and anti-gamma tubulin antibodies were used in Western blot experiments (middle and bottom panel). (C, D) 32hpf embryos injected with 1.25 pmol of (C) control morpholino (mo^C) or (D) tcf7l2 exon five splicing morpholino (mo^SPtcf7l2). Scalebar in (C) is 200 µm. (E) Plot showing estimated eye volume of 32hpf live embryos injected with injected with mo^C or mo^SPtcf7l2 as in (C, D). Error bars are mean ± SD, P values from unpaired t test. Percentage indicates average eye profile size relative to control.