IMAGE

Fig. 7

ID
ZDB-IMAGE-170517-39
Genes
Antibodies
Source
Figures for Anelli et al., 2017
Image
Figure Caption

Fig. 7

Twist expression is required for BRAFV600E mediated thyroid follicle disruption.

(A) Results from an in silico screen of transcription factor (n = 1665) expression in human PTC correlated with BRAFV600E gene signature performed by GSEA. Top hits represented in red. (B) CrisprVariants analysis of twist3 gene editing in zebrafish embryos. tg-BRAFV600E-TOM embryos were microinjected with rCas9 and either a non-targeting sgRNA or a twist3-specific sgRNA. Individual embryos were isolated and a clonal analysis of the twist3 target locus was performed. Non-targeting embryos (n = 5, grouped) had no evidence of deletions at the twist3 locus, 21/21 clones were wild-type. Twist3 targeted embryos (n = 5, one per column) had a range of small indels at the targeting site. (C) Representative confocal images of 5 dpf fixed tg-BRAFV600E-TOM embryo injected with either non-targeting or twist3 targeting sgRNA. Anti-T4 immunostaining performed to identify T4-positive (green) follicles. (D) Quantification of follicle rescue in embryos injected with a non-targeting versus twist3 targeting sgRNA in BRAFV600E-TOM embryos. Follicle rescue was observed in 0/142 non-targeting sgRNA injected embryos vs. 57/173 (32.9%) of twist3 sgRNA injected embryos, p<1.0 × 10−3, two-tailed Fisher exact test. (E) FACS plot of TdTomato-positive cells isolated from twist3 sgRNA injected tg-BRAFV600E-TOM embryos. (F) Alignment of DNA chromatogram from a clone isolated from twist3 sgRNA injected TOM+ cells harboring a three nucleotide deletion in the twist3 gene. The twist3 sgRNA sequence is underlined in red.

Figure Data
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