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. 2017 Apr 11;6:e22706. doi: 10.7554/eLife.22706

Figure 2. Knock down of mpc1 expression phenocopies loss of apc.

(A) Gross phenotype associated with mpc1 knock down. (B) Whole mount in situ hybridization analysis for organ-specific markers in mpc1 mo. Alcian blue staining revealed improper cartilage development (*) and confirmed loss of pectoral fins in mpc1 mo. pectoral fin bud (pfb), heart (h), gut (g), pancreas (p). (C) Cross section of the eye and gut from control or mpc1 mo. Prior to sectioning, embryos were previously stained with eye and gut-specific markers, irbp (red arrow) and fabp2 (black arrow), respectively. (D) Co-injection with human MPC1 RNA led to increased percentage of mpc1 mo with normal pectoral fins as determined by in situ staining for idi1. Statistical significance was analyzed using Fisher’s exact test. See also Figure 2—figure supplements 1, 2.

DOI: http://dx.doi.org/10.7554/eLife.22706.006

Figure 2.

Figure 2—figure supplement 1. mpc1 morphant phenotype is rescued by human MPC1 RNA.

Figure 2—figure supplement 1.

(A) PCR analysis amplifying mpc1 WT (322 bp) or morphant band (189 bp). Un-injected embryos (UI) served as a second control group. (B) Observed penetrance for mpc1 morpholino. (C) Observed phenotype for MPC1 mRNA overexpression in WT embryos. (D) Rescue of pectoral fins (pfb) and underdeveloped midbrain (mb) in 72 hpf mpc1 mo co-injected with human MPC1 RNA (mpc1 mo + MPC1 RNA) was determined by in situ hybridization for id1 and otx2, respectively. (E) Quantification of embryos with normal midbrain and eye development as measured by otx2 staining. Statistical significance was analyzed using Fisher’s exact test. (F) PCR analysis confirming presence of human MPC1 transcript (300 bp) in 48 hpf embryos.
DOI: http://dx.doi.org/10.7554/eLife.22706.007
Figure 2—figure supplement 2. mpc2 morphants phenocopy loss of mpc1.

Figure 2—figure supplement 2.

(A) Knockdown of mpc2 expression in WT embryos (mpc2 mo) resulted in phenotypes previously described for mpc1 morphants including enlarged hindbrain (black arrows), body deformities and absence of pectoral fins (blue arrows). (B) PCR analysis using primers specific for amplifying mpc2 WT (137 bp) or morphant band (1675 bp) confirmed knockdown of mpc2 expression in 48 hpf embryos. (C) Observed penetrance for mpc2 morpholino.
DOI: http://dx.doi.org/10.7554/eLife.22706.008