Figure 3.

Lats2 and ASPP1 interact within cells. (A) Schematic representation of full-length ASPP1 and different deletion mutants lacking either the N terminus (ΔN), the ANK repeat (ANK), and SH3 domain (ΔC), or the SH3 domain only (small ΔC). (B) HCT116 cells were transiently transfected with V5-tagged full-length ASPP1 or deletion mutants thereof, together with empty vector or myc-tagged Lats2. Lysates were immunoprecipitated (IP) with either anti-V5 or anti-myc antibodies. Westerns were immunoblotted (IB) using antibodies directed against either V5 tag, myc tag, or GAPDH. (DW) Direct Western: 2.5% of each lysate subjected directly to SDS-PAGE and Western blot analysis. (C) WI-38 cells were infected with H-RasV12 or empty retroviral vector. Two days after infection, cells were harvested and subjected to immunoprecipitation with either anti-Lats2, anti-ASPP1, or control anti-HA antibodies. Membranes were immunoblotted (IB) using anti-ASPP1, anti-Lats2 (LA2), or anti-GAPDH antibodies. Direct Western (DW) is as in B.