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. 2021 Mar 22;17(3):e1009446. doi: 10.1371/journal.pgen.1009446

Fig 8. BAF complex is essential for Brg1-Yap-Tead-dependent transcription of target genes in NCCs.

Fig 8

(A) Western blot for BAF155 and BAF170 using protein extracts from O9-1 cells transfected with control, BAF155 or BAF155/170 siRNA for 72 hours. Knockdown efficiency was evaluated by BAF155 and BAF170 western blots. Input blots for Brg1, Tead, and Yap together with β-actin controls are presented. (B) Cell extracts from control siRNA, BAF155 siRNA, and BAF155/170 siRNA transfected cells were prepared for immunoprecipitation (IP) using IgG control and anti-pan Tead antibody followed by western blotting (WB) for Brg1. IP was performed using IgG control and anti-Brg1 antibody followed by WB for Yap. Similarly, IP was performed using IgG control and anti-pan Tead antibody followed by WB for Yap. Representative blots are presented. (C) Schematic illustration of the Tead luciferase reporter construct with Tead and Yap. (D) Results of normalized luciferase reporter assays in O9-1 cells with Tead-luciferase reporter (Tead-Luc) in the presence of control siRNA or BAF155/170 siRNA. (E) Results of normalized luciferase reporter assays in HEK293T cells with Tead-Luc reporter in the presence of Brg1 alone or in combination with dominant-negative Tead1 (DN-Tead1). (F) Results of normalized luciferase reporter assays in HEK293T cells with Tead-Luc reporter in the presence of Brg1, Yap alone, or both. (G) Results of normalized luciferase reporter assays in HEK293T cells with Tead-Luc reporter in the presence of Brg1 and Yap in combination with different doses of DN-Tead1. Values are reported as means ± SD (*P < 0.05, **P < 0.01, ***P < 0.001; NS, not significant).