Fig. S4

Fig. S4

Figure S4 (related to Figure 6). Effects of oxtl and oxtlr gene knockdowns

A, Scheme depicting oxtl gene structure indicating the binding site of a splice blocking antisense oligonucleotide (oxtl-MO).

B, Reduction of Oxytocin (Oxtl) protein expression by oxtl-directed antisense oligonucleotide (oxtl-MO). Anti-oxytocin immunostaining of the neurohypophysis of 3-day old transgenic oxtl:EGFP embryos. Injection of oxtl-MO causes a marked reduction in Oxtl but not EGFP protein levels.

C, Scheme depicting oxtlr gene structure indicating the binding site of a splice blocking antisense oligonucleotide (oxtlr-MO) as well as the PCR primers used to amplify the respective exons (Ex) of the various mRNA products. A table summarizing the expected mRNA sizes in the control and followed oxtlr knockdown is shown at the bottom.

D, Gel electrophoresis analysis of PCR-amplified mRNA species in control and in embryos injected with oxtlr-MO. Primer pairs used to amplify the respective exons (Ex) are indicated at the top. Amplification of exon 1 (Ex1F/Ex1R) was used as control to demonstrate untargeted constitutive exon, which was not affected by the knockdown reagent.

E, Scatterplot representation of the fluorescence profiles of cell suspensions from dissociated 3-day old vegfr2:mCherry embryos (right) and their non-transgenic siblings (left), prior to FACS sorting. P1 is the proportion of cells that exhibit high red fluorescence (defined as ‘mCherry+’ cells, ~1% of the total population). P4 is the proportion of cells that exhibit no fluorescence (defined as ‘mCherry-’ cells, ~80% of the total population).