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. 2000 Mar;11(3):999–1010. doi: 10.1091/mbc.11.3.999

Figure 4.

Figure 4

hStau and L22 interact with telomerase and hTERT. (A) hStau and L22 associate with telomerase activity in 293 cells. Telomerase assays (modified TRAP, see MATERIALS AND METHODS) were performed on a 293 extract (assayed at 5 μg of protein; lane1) and immunoprecipitation pellets (lanes 2–13) from immunoprecipitations with various antibodies. The antibodies used are indicated at the top of the gel. Lanes 2 and 3, precipitation using hStau preimmune serum; lanes 4 and 5, precipitation using anti-hStau antibody; lanes 6 and 7, precipitation using anti-GST antibody; lanes 8 and 9, precipitation using L22 preimmune serum; lanes 10 and 11, precipitation using anti-L22 antibody; and lanes 12 and 13, precipitation using anti-L22 antibody preincubated with L22 peptides. Samples in lanes 3, 5, 7, 9, 11, and 13 were pretreated with RNase (+) before the telomerase reactions. (B) hStau and L22 interact with hTERT. Western blot analysis using anti-HA (top), anti-hStau (middle), or anti-L22 (bottom) antibody on cell lysates (lanes 1 and 2) or various immunoprecipitation pellets (lanes 3–8) is shown. Extracts used are a mock-transfected 293 extract (lanes 1, 3, and 6) and an hTERT-HA–transfected cell extract (lanes 2, 4, 5, 7, and 8). The antibodies used in immunoprecipitation reactions were the following: lanes 3 and 5, precipitation using anti-L22; lane 4, precipitation using L22 preimmune serum; lanes 6 and 8, precipitation using anti-hStau; and lane 7, precipitation using hStau preimmune serum. The relative mobility of molecular weight markers (in kilodaltons) is indicated on the left. The identity of various bands is indicated by arrows.