FIGURE

Fig. 1

ID
ZDB-FIG-120417-1
Publication
Tallafuss et al., 2012 - Turning gene function ON and OFF using sense and antisense photo-morpholinos in zebrafish
Other Figures
All Figure Page
Back to All Figure Page
Fig. 1

S-photo-MO and AS-photo-MO regulate Ntla function. (A,B) Diagrams describing the mechanism of AS-photo-MO (A) and S-photo-MOs (B). (C) Molecular structure of the photo-subunit (red) and its position within the MO (green). (D,E) Lateral view of a wild-type embryo (D) and an ntla-MO injected embryo, missing posterior trunk and tail (E) at 48 hpf. Scale bar: 250 μm. (F-I) Overlays showing bright field and anti-Ntla labeling (green) in injected embryos at 26-28 hpf. Examples of severe (F), medium (G), mild (H) and normal (I) phenotypes. (J-L) Graphs showing percentage of embryos injected with MOs and photo-MOs resulting in severe (red), medium (orange), mild (light blue) and normal (dark blue) phenotypes, based on morphology. (J) ntla-S-photo-MOs of a length of 21 nucleotides (S21-photo-MO) and/or 25 nucleotides with four mismatches (S25-photo-MO) allow or block protein formation dependent on UV exposure. MO, n=60; MO UV, n=179; MO+S21-photo-MO, n=58; MO+S21-photo-MO UV, n=125; MO+S25-photo-MO, n=61; MO+S25-photo-MO UV, n=116; WT, n=114; WT UV, n=135. (K) All ntla-AS-photo-MO tested sufficiently block ntla gene function. Introducing two mismatches on either side of the photo-subunit (AS-4mis-photo-MO) significantly improved dissociation rate from the target and therefore recovery (P<0.05) of the phenotype after photo-cleavage compared with AS-photo-MO. AS-photo-MO, n=84; AS=photo=MO UV, n=132; AS-2mis-photo-MO, n=113; AS-2mis-photo-MO UV, n=134; AS-4mis-photo-MO, n=112; AS-4mis-photo-MO UV, n=152. Asterisks indicate significant differences (P<0.05) between untreated and UV-treated embryos. Student’s t-test includes data in groups ‘severe’ and ‘normal’. (L) Distribution of phenotypes after injecting different ratios of ntla-MO to S25-photo-MO solutions. The horizontal bar marks samples exposed to UV. (M) Quantification of in vitro mass spectroscopy analyses showing percentage of intensity of active (intact photo-element, red, pink) and inactive (cleaved photo-element, blue) ntla- or gfp-AS-photo-MOs, MO1 and MO2 respectively, measured over time after UV exposure. (N,O) Embryos injected with gfp-AS-photo-MO without UV exposure (N) and with UV exposure (O). Scale bar: 30 μm.

Expression Data

Expression Detail
Antibody Labeling
Phenotype Data

Phenotype Detail
Acknowledgments
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Development