FIG 6.

Effects of calyculin A and proteasome inhibitors on TcRNA Pol II phosphorylation. (A) Control cells or cells incubated with 50 nM calyculin A for 2 h were processed for Western blotting using total lysates. The chromatin-associated (Chr) and soluble (Sol) fractions were probed with anti-TcRNA Pol II antibodies. (B) Immunofluorescence labeling with anti-TcRNA Pol II antibodies and DAPI staining of control or calyculin A-treated cells (k, kinetoplast; N, nucleus). Bar = 2 μm. (C) Amount of TcRNA Pol II labeling (means ± standard deviations; n = 30) associated with the SL relative to the other areas of the nucleus in control and calyculin-treated samples. (D) Cells were treated with 10 μM MG132 and 10 μM lactacystin for 2 h, and the total, chromatin-associated, and soluble fractions were analyzed by Western blotting using anti-TcRNA Pol II.