A. Immunoblot of lysates prepared from cells transfected with plasmids encoding FLAG-tagged Rax N-terminal mutations using antibodies raised against the FLAG epitope tag. Lanes: 1 – empty vector; 2 – Rax FL; 3 – L38P; 4 - ΔOP; 5 – ΔN31; 6 – ΔN39. B. Immunoblot of lysates prepared from cells transfected with plasmids encoding FLAG-tagged Rax C-terminal truncations using antibodies raised against the FLAG epitope tag. In this blot we observed a nonspecific band of approximately 60 kDa. This band was not observed in all blots. Lanes: 1 – empty vector; 2 – Rax FL; 3 - ΔC293; 4 – ΔC277; 5 – ΔC244; 6 – ΔC212. C. Immunoblot of lysates prepared from cells transfected with plasmids encoding FLAG-tagged Rax internal deletions and point mutations using antibodies raised against the FLAG epitope tag. Lanes: 1 – empty vector; 2 – Rax FL; 3 – ΔRX; 4 – ΔPPXY; 5 – 3PA. D. Immunoblot of lysates prepared from cells transfected with plasmids encoding FLAG-tagged Rax N-terminal mutations and C-terminal truncations using antibodies raised against the FLAG epitope tag. Lanes: 1 – Rax FL; 2 – L38P + ΔC293; 3 – ΔOP + ΔC293; 4 – L38P + ΔC244; 5 – ΔOP + ΔC244. E. Immunoblot of lysates prepared from cells transfected with GAL4 DBD fusion proteins using antibody raised to GAL4 DBD. Lanes: 1 – GAL4 DBD; 2 – GAL4 DBD+enR; 3 – GAL4 DBD+VP16; 4 – GAL4 DBD+Rax C290–322; 5 – GAL DBD+Rax273–322; 6 – GAL4 DBD+Rax C245–322; 7 – GAL4 DBD+Rax C213–322. F. Immunoblot of lysates prepared from cells transfected with FLAG-tagged Rax and Rax2 using antibodies raised against the FLAG epitope tag. Lanes: 1 – Rax FL; 2 – Rax2 FL; 3 – Rax2 ΔC180. In all panels, numbers at the left edge of the blot represent sizes of molecular weight markers (kDa). To access the data, click or select the words “Appendix 1.”