ZFIN Image: Ahmed et al., 2022, Figure 1

Image

Figure Caption

Figure 1—figure supplement 1

Retinal lamination defects in strip1^rw147 mutants are phenocopied by either CRISPR- or MO-mediated knockdown of strip1 and rescued by overexpression of Strip1.

(A) CRISPR-mediated, targeted mutagenesis of zebrafish strip1 gene. A 10-bp deletion was introduced into exon 9, leading to a premature stop codon in strip1 gene. (B) Morphology of wild-type and strip1^crispr^Δ¹⁰ embryos at 4 dpf. Dotted lines demarcate the eye outline. The arrowhead indicates an abnormal lower jaw. An asterisk indicates heart edema. (C) Plastic sections of wild-type and strip1^crispr^Δ¹⁰ mutant retinas stained with toluidine blue at 4 dpf. (D) A polyacrylamide gel image showing PCR amplicons using primers that flank the 10-base deletion of strip1^crisprΔ10. Wild-type and strip1^crisprΔ10 mutant bands are expected to appear at 279 and 269 bps, respectively. Different band sizes were consistently obtained from progeny produced by pairwise crosses of strip1^crisprΔ10 heterozygous mutant fish. (E) Morphology of non-injected and MO-strip1-injected wild-type embryos at 4 dpf. Dotted lines outline the eye. An arrowhead indicates the abnormal lower jaw. An asterisk indicates heart edema. (F) Retinal plastic sections of non-injected and MO-strip1-injected wild-type embryos at 4 dpf. (G) Western blotting of 2-dpf head lysates from non-injected and MO-strip1-injected wild-type embryos. Red and black arrows show expected band sizes for zebrafish Strip1 (93 kDa) and the loading control β-actin (42 kDa), respectively. (H) Plastic sections of 4 dpf wild-type and strip1^rw147 mutant retinas without transgene (top panels), and with the transgenes Tg[hsp:WT.Strip1-GFP] (middle panels) or Tg[hsp:Mut.Strip1-GFP] (bottom panels) post heat shock treatment. Overexpression of Green Fluorescent Protein (GFP)-tagged wild-type Strip1 rescues retinal defects in strip1^rw147 mutants, while the GFP-tagged mutant form fails to rescue. (I) A staggered bar graph, which represents the percentage of strip1^rw147 mutants with a defective retina vs. mutants with a rescued retina in strip1^rw147 mutants without transgenes, and with Tg[hsp:WT.Strip1-GFP] or Tg [hsp:Mut.Strip1-GFP].

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 @ Elife

Figure 1—figure supplement 1

Figure 1—figure supplement 1

Retinal lamination defects in strip1rw147 mutants are phenocopied by either CRISPR- or MO-mediated knockdown of strip1 and rescued by overexpression of Strip1.

Retinal lamination defects in strip1^rw147 mutants are phenocopied by either CRISPR- or MO-mediated knockdown of strip1 and rescued by overexpression of Strip1.