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Fig. S8. Genetic interaction of sp8 and pax2, pax8, and fgf8 was evaluated by oc90 expression in single-injected or coinjected embryos. No strong interactions or synergies were observed. The number of analyzed embryos is in parentheses. Serial dilution of each factor determined the lowest dose of mRNA, which induced ectopic otic tissues. Microinjection of pax2, pax8, or fgf8 alone resulted in the development of OVs that were reduced or enlarged in size, but did not induce ectopic otic tissues. A higher concentration of pax2 has a potential to produce ectopic OV, which partially agrees with previous studies (1). When 12.5 pg of pax2 was coinjected with sp8 mRNA (12.5 pg), there was an approximately fourfold increase in the number of embryos that show ectopic oc90 expression domain, compared with a single injection of 12.5 pg of either sp8 or pax2 (which usually led to otic vesicle reduction). At higher pax2/sp8 combined con- centrations, however, the domain of oc90 expression more frequently developed enlarged otic tissues. Unlike pax2/sp8 coinjection, 7% of pax8/sp8-coinjected embryos showed ectopic otic tissues and this is similar to sp8 (6%) single injections. Pax8/sp8 coinjection resulted in reduction of oc90 expression in 33% of embryos, which is higher than sp8 (18%) and pax8 (12%) single injections. Fgf8/sp8 coinjection most frequently reduced oc90 expression, more frequently than single injections of sp8 and fgf8, 18% and 15%, respectively. Image published in: Chung HA et al. (2014) Copyright © 2014. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License. Larger Image Printer Friendly View

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