Figure S1. RNAi depletion efficiencies.... | Rockefeller University Press

Figure S1.

$RNAi depletion efficiencies. (A) Immunoblot on the cell lysate of U2OS cells expressing an siRNA-resistant GTSE1-GFP BAC transgene and transfected with control (Ctrl) or CHC siRNA for 66 h. Cells were concomitantly transfected with GTSE1 siRNA to deplete endogenous GTSE1. Immunoblots are with anti-GTSE1, anti-CHC, or anti-α-tubulin antibodies. Anti-CHC blot was performed on same samples run on a different gel. (B) Quantification of CHC and GTSE1-GFP on immunoblot of three siRNA experiments performed as in A. GTSE1-GFP and CHC levels were normalized to α-tubulin and presented in percentage of the control siRNA. Mean percentages (± standard error) are presented (n = 3 experiments). (C) Immunoblot on the cell lysate of mESCs (R1\E) transfected with control, mGTSE1, CHC, or mGTSE1 + CHC siRNA. Immunoblots are with anti-CHC, anti-GTSE1, and anti-α-tubulin antibodies. (D) Immunoblot on the cell lysate of U2OS cells stably expressing a CHC-GFP BAC transgene and transfected with control or GTSE1 siRNA. The immunoblot is with anti-GTSE1 and anti-α-tubulin antibodies. (E) Immunoblot on asynchronous cell lysates of GTSE15xLID cells transfected with control, GTSE1, MCAK, or GTSE1 + MCAK siRNA. Immunoblots are with anti-GTSE1, anti-α-tubulin, or anti-MCAK. (F) Immunoblot on asynchronous cell lysates of U2OS cells transfected with control, CHC, GTSE1, MCAK, CHC + GTSE1, or CHC + MCAK siRNA. Immunoblots are with anti-CHC, anti-GTSE1, anti-MCAK, or anti-α-tubulin.$

RNAi depletion efficiencies. (A) Immunoblot on the cell lysate of U2OS cells expressing an siRNA-resistant GTSE1-GFP BAC transgene and transfected with control (Ctrl) or CHC siRNA for 66 h. Cells were concomitantly transfected with GTSE1 siRNA to deplete endogenous GTSE1. Immunoblots are with anti-GTSE1, anti-CHC, or anti-α-tubulin antibodies. Anti-CHC blot was performed on same samples run on a different gel. (B) Quantification of CHC and GTSE1-GFP on immunoblot of three siRNA experiments performed as in A. GTSE1-GFP and CHC levels were normalized to α-tubulin and presented in percentage of the control siRNA. Mean percentages (± standard error) are presented (n = 3 experiments). (C) Immunoblot on the cell lysate of mESCs (R1\E) transfected with control, mGTSE1, CHC, or mGTSE1 + CHC siRNA. Immunoblots are with anti-CHC, anti-GTSE1, and anti-α-tubulin antibodies. (D) Immunoblot on the cell lysate of U2OS cells stably expressing a CHC-GFP BAC transgene and transfected with control or GTSE1 siRNA. The immunoblot is with anti-GTSE1 and anti-α-tubulin antibodies. (E) Immunoblot on asynchronous cell lysates of GTSE1^5xLID cells transfected with control, GTSE1, MCAK, or GTSE1 + MCAK siRNA. Immunoblots are with anti-GTSE1, anti-α-tubulin, or anti-MCAK. (F) Immunoblot on asynchronous cell lysates of U2OS cells transfected with control, CHC, GTSE1, MCAK, CHC + GTSE1, or CHC + MCAK siRNA. Immunoblots are with anti-CHC, anti-GTSE1, anti-MCAK, or anti-α-tubulin.