Plasmids constructed for Y2H and HEK293T cell transfection assay.

(A) Full length and truncated MTL5 were cloned into GAL4 activation domain (AD) vector pGADT7 to generate AD-MTL5, AD-MTL5-N (1–250 aa), AD-MTL5-M (251–370 aa), AD-MTL5-C (371–475 aa), AD-MTL5-C1 (371–418 aa), AD-MTL5-C2 (419–442 aa) or AD-MTL5-C3 (443–475 aa). (B) Full length and truncated LIN9 were cloned into GAL4 BD/bait vector pGBKT7 to generate BD-LIN9, BD-LIN9-N (1–150 aa), BD-LIN9-M (151–340 aa) or BD-LIN9-C (341–559 aa). (C) The fragments of MTL5-N (1–250 aa), MTL5-M (251–370 aa), MTL5-C1 (371–418 aa), MTL5-C2 (419–442 aa) or MTL5-C3 (443–475 aa) were deleted from MYC-Mtl5 to generate MYC-Mtl5-ΔN, MYC-Mtl5-ΔM, MYC-Mtl5-ΔC1, MYC-Mtl5-ΔC2, or MYC-Mtl5-ΔC3, respectively. The gray boxes represent the cysteine-rich domains (CRC). The green and red boxes represent the regions containing helix domains predicted by SWISS-MODEL. del, deletion;aa, amino acid.

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