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. 2024 Jul 31;19(7):e0293852. doi: 10.1371/journal.pone.0293852

Fig 8. Effects of Zbtb11 on the interaction between Otx2 and Tle1.

Fig 8

(A,B) Co-immunoprecipitation (Co-IP) assays and western blots were performed using injected embryos as described in the Fig 7 legend. (A) No specific interaction between Zbtb11 and Tle1. mRNA for Venus-Zbtb11 was co-injected with or without mRNA for HA-Tle1 as indicated. (B) Enhancement of the interaction between Otx2 and Tle1 by Zbtb11. mRNA for HA-Tle1 was co-injected with combinations of mRNA for Myc-Otx2, Venus-Zbtb11, or Venus-NLS, as indicated. The band of HA-Tle1 co-immunoprecipitated with Myc-Otx2 is increased by co-expression of Venus-Zbtb11 (lane 2), compared to that of Venus-NLS as a control (lane 5). Black arrowheads, nascent products (A,B). Asterisks indicate non-specific bands (A). Orange arrowheads, co-immunoprecipitated HA-Tle1; blue brackets, modified Myc-Otx2; black dots, IgG heavy chains from the anti-Myc antibody (B). (C) The effect of Zbtb11 on transactivation activity of Myc-Otx2 analyzed by luciferase reporter assays. mRNA for Zbtb11 with combinations of mRNA for Myc-Otx2 was co-injected with SOP-FLASH reporter DNA, as indicated. Amounts of injected mRNAs (pg/embryo): Myc-otx2, 100; zbtb11, 1500. (D) The effect of Zbtb11 on transrepression activity of Otx2 and Tle1 analyzed by luciferase reporter assays. mRNA for Zbtb11 with combinations of mRNA for Otx2 and Tle1 was co-injected with the meis3-D2-luc reporter DNA, as indicated. Amounts of injected mRNAs (pg/embryo): otx2, 20; tle1, 20; zbtb11, 1500. *P < 0.05, **P < 0.01 (t-test); error bars, standard error of the mean (s.e.m.); NS, not significant; n, the total number of samples (C,D).