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. 2010 May 3;189(3):527–539. doi: 10.1083/jcb.200912125

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© 2010 Etard et al.

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Figure 3. — Apo2a morphants have defects in the skeletal musculature and the heart. (A) Position of the morpholinos Mo(ATG)-apo2a (ATG) and Mo(UTR)-apo2a (UTR) on the apo2a mRNA. (B–D) Mo(ATG)-apo2a (B and D) and control Mo(ATG)-apo2a-cont (harboring five mismatches relative to the Mo(ATG)-apo2a sequence; C)-injected embryos. The morphants exhibit a curved body axis (B) and show reduced birefringence (D) compared with controls (C). Note that the Mo(ATG)-apo2a–injected embryos (B and D) were coinjected with the Mo-p53 to suppress unspecific effects (see K–M for additional controls for Mo-p53 injection). (E–G) Mo(UTR)-apo2a (E and G)- and control Mo(ATG)-apo2a-cont (F)-injected embryos. The morphants exhibit a curved body axis (E) and show reduced birefringence (G) compared with controls (F). (H–J) Apo2a-GFP expression (H) in embryos injected with an apo2a-gfp encoding plasmid. Co-injection of Mo(ATG)-apo2a with the apo2a-gfp plasmid abolishes Apo2a-GFP expression (I), whereas coinjection of the apo2a-gfp plasmid with the five-mismatch Mo(ATG)-apo2a-cont does not abolish Apo2a-GFP expression (J). (K–M) Uninjected (K), Mo(ATG)-apo2a-cont– and Mo-p53–coinjected (L), and Mo(ATG)-apo2a– and Mo-p53–coinjected embryos (M). Injection of Mo(ATG)-apo2a morpholino reduces birefringence (M), whereas neither the five-mismatch Mo(ATG)-apo2a-cont nor the Mo-p53 had an effect on the birefringence of the musculature (L). (N) Mo(ATG)-apo2a and Mo(UTR)-apo2a morphants have a decreased heart rate (the graph shows heart beats per minute, n = 20 embryos). Black bars represent standard deviation. The datasets were pairwise subjected to the Student’s t test. The difference between the heart rate of Mo(ATG)-apo2a versus Mo(UTR)-apo2a was not significant (P = 0.220), whereas the difference between the heart rate of Mo(ATG)-apo2a versus control or Mo(UTR)-apo2a versus control was highly significant (P = 0.002 and P = 0.003, respectively). (O) Pericardial edema (arrow) in a Mo(ATG)-apo2a– and Mo-p53–coinjected embryo. Embryos shown are 72 hpf, except F–H, which show 24-hpf embryos. Pigmentation of the embryo shown in E was suppressed by 1-phenyl 2-thiourea (PTU) treatment (Karlsson et al., 2001). Bars, 80 µm.