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Figure 5. Examples on identification and de-convolution of active regulators.(A–B) identification of the MAPK/Erk activator fbxo43 (erp1). (A) Western blot of stage 8 embryos injected with 12 pools (01–12) derived from plate #01 and 7 pools (01–07) from plate #02 and probed with anti-phospho-Erk (pErk) antibody. The arrow indicates increased Erk phosphorylation upon injection of mRNAs derived from plate #02, pool 07. (B) De-convolution of the above pool. Embryos injected with single RNAs were collected at stage 8 and uninjected lysate from stage 8 and stage 10.5 were used as negative and positive control of Erk phosphorylation respectively. The arrow indicates the active clone of TEgg009F05, identified in plate #2, column 08, row G. This clone was confirmed as the X. tropicalis fbxo43 (erp1) gene. (C–D) Identification of PI3K/Akt inhibitor prkaca. (C) Western blot of stage 10.5 embryos injected with 24 pools (01–12) derived from plate #09 and #10 and probed with anti-phospho-Akt (pAkt) antibody. The arrow indicates decreased Akt phosphorylation upon injection of mRNAs derived from plate #10, pool 02. (D) De-convolution of the above pool. mRNA synthesis, injection, and Western blot were performed as in (B) except that stage 10.5 embryos were used. The arrow indicates the active clone of TEgg046d13 is identified in plate #09, column 02, row E. This clone was later identified as encoding the X. tropicalis prkaca gene. Image published in: Zhang S et al. (2013) Image reproduced on Xenbase with permission of the publisher and the copyright holder. This image is reproduced with permission of the journal 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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