Figure Caption
Molecular and phenotypic characterization of rb1off and rb1on alleles.(A) Diagram of the rb1off allele. (B) Plot of RT-qPCR results from wild type +/+ (n=3), heterozygous rb1off/+ (n=3), and homozygous rb1off/off (n=3) larvae showing the relative level of rb1 mRNA transcript using reference gene rps6kb1b. Primer pairs were located in exons 6 and 8, or downstream exons 23 and 24. (C – E) Gross phenotype of wildtype +/+ (C), rb1off/stop (D), and rb1Δ7/stop (E) 5 dpf larvae. Arrowhead in D points to rb1off allele gcry1:BFP secondary reporter expression in lens. Asterisk marks the rb1stop allele mly7:GFP secondary reporter expression in heart. (F – H) pH3 and HuC/D labeling of sectioned head tissue from 5 dpf +/+ (F), rb1off/stop (G), and rb1Δ7/stop (H). (I) Diagram of the rb1on allele. (J) Plot of RT-qPCR results from wild type +/+ (n=3), heterozygous rb1on/+ (n=3), and homozygous rb1on/on (n=3) larvae showing the relative level of rb1 mRNA transcript using reference gene rps6kb1b. Primer pairs were located in exons 6 and 8, or downstream exons 23 and 24. (K, L) Gross phenotype of wildtype +/+ (K), heterozygous rb1on/+ (L) and transheterozygous rb1on/stop (M) 5 dpf larvae. Arrowhead in L, M points to rb1off allele gcry1:BFP secondary reporter expression in lens. Asterisk in M marks the rb1stop allele mly7:GFP secondary reporter expression in heart. pH3 and HuC/D labeling of sectioned head tissue from 5 dpf +/+ (N), heterozygous rb1on/+ (O) and transheterozygous rb1on/stop (P).OT, optic tectum; R, retina; Th, thalamic region. Error bars represent mean ± s.e.m. Scale bars: 200 μm (C–E, K–M), 50 μm (F–H, N–P).
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