PMC

Depletion of chordin mRNA in Xenopus embryos caused defects in pronephros and muscle formation. A: Blot hybridization of RNA isolated from uninjected control embryos (lane 1), embryos injected with increasing amounts of the control N,N-diethyl-ethylenediamine (DEED) oligonucleotide (lanes 2-4), and embryos injected with increasing amounts of the chordin DEED-antisense oligonucleotide (lanes 5-7). The blot was hybridized with probes to detect chordin and goosecoid mRNA. B-E: Morphology of chordin mRNA-depleted embryos. B: Embryos injected with chordin-antisense DEED oligo. C: Embryo injected with control DEED oligonucleotide. D: Uninjected control embryo. E: Embryo injected with chordin mRNA + chordin-antisense DEED oligo. F-H: Analysis of pronephric duct formation in chordin-depleted embryos using immunocytochemistry with the 4A6 antibody. F: Injected with chordin-antisense DEED oligo. G: Injected with control DEED oligo. H: Uninjected control. I-K: Analysis of pronephric tubule formation in chordin-depleted embryos using immunocytochemistry with the 3G8 antibody. I: Injected with chordin-antisense DEED oligo. J: Injected with control DEED oligo. K: Uninjected control. L-N: Analysis of somitic muscle formation in chordin-depleted embryos using immunocytochemistry with the 12-101 antibody. L: Uninjected control. M: Injected with control DEED oligo. N: Injected with chordin-antisense DEED oligo.

Reduction of XNkx2-10 leads to abnormal development. (A–G) Pictures of stage 22 embryos. Scale bar is 3 mm. (A) are non-injected embryos. (B) Embryos injected with 2 ng of XNkx2-10 mRNA (overexpression). (C) Morpholino control injected embryos. (D) Morpholino injected embryos. (E) Deed oligonucleotide injected embryos. (F) Embryos injected with both morpholino and 2 ng of XNkx2-10 mRNA (morpholino rescue). (G) Embryos injected with both Deed oligonucleotide and 2 ng of XNkx2-10 mRNA (Deed rescue), (a1) Stage 42 non-injected embryo. Scale bar is 3 mm. (b1) Stage 42 overexpression embryo, (c1) Stage 42 morpholino control injected embryo, (d1) Morpholino injected embryo. It has an abnormal head shape and delayed eye development (white arrows). (d2) Morpholino injected embryo that is stunted, has an abnormal head shape and delayed eye development, (e1) Deed oligonucleotide injected embryo that is stunted, has an abnormal head shape and delayed eye development. (e2) Deed injected embryo that is stunted, has an abnormal head shape, delayed eye development and delayed gut development, (f1) Stage 42 morpholino rescue embryo, (g1) Stage 42 Deed rescue embryo. (H) Western blot demonstrating that injection of either the morpholino or Deed oligonucleotide leads to reduced XNkx2-10 protein levels in stage 22 embryos (black arrows) and a Western blot demonstrating the translation of rescue mRNA (T7) in stage 22 embryos. (I) Western blot demonstrating XNkx2-10 and nucleolin protein levels at stage 36. (J) Western blot demonstrating XNkx2-10 and nucleolin protein levels at stage 46. Nucleolin serves as a positive control. The expected size of XNkx2-10 is 29.2 kDa while the expected size of nucleolin is 95 kDa. Lane 1 is protein from non-injected embryos. Lane 2 is protein from morpholino control injected embryos. Lane 3 protein from morpholino injected embryos. Lane 4 contains protein from Deed oligonucleotide injected embryos. Lane 5 contains protein from Deed rescue embryos.

(A) A 6F sheath introduced next to the Impella catheter into the 14F Impella sheath (Abiomed, Danvers, MA). (B) Filling 2 needs with 1 deed. Percutaneous coronary intervention (PCI) through the Impella sheath system.

Box plot demonstrating the range of maximum ventricular area values. Sample 1 contains data from non-injected embryos. Sample 2 contains data from morpholino control injected embryos. Sample 3 contains data from XNkx2-10 overexpression embryos. Sample 4 contains data from XNkx2-10 morpholino injected embryos. Sample 5 contains data from XNkx2-10 Deed oligonucleotide injected embryos. Sample 6 contains data from morpholino rescue embryos. Sample 7 contains data from Deed rescue embryos. The morpholino and Deed oligonucleotide significantly reduced the maximum ventricular area of their respective hearts. Alternatively, co-injection of either the morpholino or Deed oligonucleotide with 2 ng of XNkx2-10 mRNA (rescue) restored the size of the maximum ventricular areas. The * symbol indicates a p value of less than 0.00001.