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Fig. S4

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ZDB-IMAGE-121213-3
Source
Figures for Fior et al., 2012
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Figure Caption

Fig. S4 Generation and validation of the hsp70:HA-msgn1 transgenic line. (A) The construct used to generate the hsp70:HA-msgn1 transgenic line. The N-terminus of the msgn1 gene was fused with an HA tag and placed under an hsp70 promoter. In addition, the b-crystallin promoter was used to drive CFP in the lens to facilitate identification of transgenic embryos. (B-N′) Embryos were obtained from a cross between hsp70:HA-msgn1 heterozygous and wt fish, generating a batch with an expected frequency of 50% transgenics and 50% wt control siblings. (B-E′) In situ hybridisation showing msgn1 mRNA levels in hsp70:HA-msgn1 transgenic embryos and their wt sibling controls, with no heat shock (B) or heat shocked for 1 hour at the 13-somite stage and fixed immediately (C,C′), 2 hpHS (D,D′) and 7 hpHS (E,E′). (F-K′) Levels of HA-tagged Msgn1 protein (FH′) at the level of the tenth somite in hsp70:HA-msgn1 transgenic embryos fixed immediately after heat shock (0 hpHS), 2 hpHS and 7 hpHS and their corresponding control siblings. (I-K′) The same embryos as in F-H′ counterstained with DAPI to reveal the nuclei. (L-N′) Expression of ntl, wnt8 and tbx24 in hsp70:HA-msgn1 transgenic embryos and their respective control siblings heat shocked for 1 hour at the 7-somite stage and fixed 7 hpHS. Red arrowhead, ectopic expression of tbx24 in the tailbud. hpHS, hours post-heat shock.

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