Figure 2.

Validating effect of intron based RNAi approach on endogenous miRNA processing. (A–C), (E–G) and (I–K) Ubiquitously expressing mCherry fluorescent embryos from Tg(ubi:mCherry:SPmiR137) incrosses were injected with 25 pg of Tol2 transposase combined with 50 pg of control transgenes 157-control or anti-miR137 expressing constructs—153 (without intron), 155 (with intron). All injected plasmids led to mosaic eGFP expression. Plasmids 153 and 155, but not 157-control, led to a reduction of red fluorescence, hence knockdown of the integrated mCherry sensor transgene. A minimum of six larvae were analyzed per condition to estimate knockdown efficiency. Graphs D, H and L are representations of pixel intensities (each dot presents a grey value) of green versus red fluorescence (presented as a percentage) from z-stacks imaged with a confocal microscope at 4 day-post-fertilization (dpf). Scale bar is 100 μm.