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Fig. 5
Foxo3b represses the transcriptional activity of Gata1 and Spi1 through protein–protein interaction. (A) Gata1 transcriptional activity was significantly inhibited by over-expression of foxo3b. (B) Spi1 transcriptional activity was significantly inhibited by over-expression of foxo3b. (C) The suppression of Gata1 transcriptional activity was confirmed by gata1 reporter assays. (D) The suppression of Spi1 transcriptional activity by foxo3b was confirmed by spi1 reporter assays. The dosages of injected plasmids: gata1-reporter, spi1-reporter, 28.0 pg/per embryo; HA-empty, HA-gata1, HA-spi1, HA-foxo3b, HA-myoD, 37.5 pg/embryo; pFR, 25.0 pg/per embryo; pRL-TK, 25.0 pg/per embryo. (E) GFP-tagged foxo3b (GFP-foxo3b) co-localized with RFP-tagged gata1 (RFP-gata1) in COS-7 cells. (F) GFP-tagged foxo3b (GFP-foxo3b) co-localized with RFP-tagged spi1 (RFP-spi1) in COS-7 cells. (G) Foxo3b interacted with Gata1 in 293 T cells as revealed by co-immunoprecipitation assays. (H) Foxo3b interacted with Spi1 in 293 T cells as revealed by co-immunoprecipitation assays. IB, immunoblot; IP, immunoprecipitation; TCL, total cell lysate.
Reprinted from Developmental Biology, 388(1), Hu, B., Zhang, W., Feng, X., Ji, W., Xie, X., and Xiao, W., Zebrafish eaf1 suppresses foxo3b expression to modulate transcriptional activity of gata1 and spi1 in primitive hematopoiesis, 81-93, Copyright (2014) with permission from Elsevier. Full text @ Dev. Biol.