Fig. 4

Expression of evx1, but not cx43, is altered in transgenic fish following overexpression of exogenous Xl-kcna5. A) Overview of experimental design. At day 0, fins from non-transgenic and transgenic siblings were amputated at the 50 % level and were systemically heat shocked at 37 °C for 1 h at 3 dpa. Fins were harvested and fixed at 4 dpa/1 dpt for cx43 or at 87 hpa for evx1 and processed for ISH or qPCR. B) Relative cx43 gene expression is shown. Amputation planes are denoted by the white dotted line. C) Gene expression levels for cx43 were quantified via qPCR. Graph shows each biological replicate (comprised of the average of two technical replicates for each sample), mean fold difference, and standard deviation. Expression level of cx43 shows no significant difference between sibling-paired negative and positive fish (unpaired t-test, p-value = 0.26). D) Fins were harvested and fixed for ISH for evx1. Amputation planes are denoted by the white dotted line. Plus (+) or minus (−) signs denote scoring of each ray for evx1 expression. E) Quantification of percentage of rays expressing evx1 shows significant decrease in evx1 positive rays following heat shock fish (unpaired t-test, p-value <0.0001). Three independent trials were conducted. dpa, days post-amputation; dpt, days post-treatment; hpa, hours post-amputation; hpt; hours post-treatment; ISH, in situ hybridization; qPCR, quantitative RT-PCR.