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. 2001 Jun 1;15(11):1383–1392. doi: 10.1101/gad.901101

Figure 5.

Figure 5

Interactions between TSC1 and TSC2. (A–C) Wing margins containing TSC1− (A), TSC2− (B), and TSC1− TSC2− (C) clones, respectively. Note that TSC single- or double-mutant bristles (marked by y−, indicated by a line above the wing margin) are similar in size. (D) Schematic representation of epitope-tagged TSC1 and TSC2 proteins that were expressed in S2 cells. The TSC1 construct corresponds to the full-length protein and migrates as an ∼150-kD protein. Three TSC2 constructs were generated, including the full-length (TSC2, >200 kD), N-terminal half (TSC2N, ∼130 kD), and C-terminal half (TSC2C, ∼130 kD). (E) S2 cells expressing TSC1/TSC2, TSC1/TSC2N, and TSC1/TSC2C constructs were lysed and total cell lysate was immunoprecipitated (IP) with α-MYC and immunoblotted with α-V5 (left panel). TSC2 or TSC2N, but not TSC2C, can be immunoprecipitated with TSC1. The same blot was stripped and reprobed with α-MYC to show that TSC1 was expressed at comparable levels in all lanes (middle panel). As an additional control, TSC2, TSC2N, and TSC2C were precipitated from the same cell lysate by using Ni-NTA agarose (α-His) and probed with α-V5. These proteins were expressed at comparable levels (right panel). (F–H) Wings expressing MS1096/UAS-TSC1 (F), MS1096/UAS-TSC2 (G), and MS1096/UAS-TSC1; UAS-TSC2, respectively. These images were taken under the same magnification. Wings overexpressing both TSC1 and TSC2 are ∼28% the size of wings expressing TSC1 or TSC2 alone.