Figure S2.

LIC1 C-terminal phosphomutants cannot rescue LIC1 depletion–induced mitotic arrest. (A) Immunoblots depicting the expression of endogenous LIC1/stably expressing hLIC1-MTAP constructs under the indicated conditions. (B) Epifluorescence micrograph of U2OS cells stained with the viable DNA dye Syto 13 (green) to visualize mitotic cells (arrows; zoomed inset shows metaphase cell). Scale bar = 50 µm. (C) Mitotic index measurements (fraction of mitotic cells) under the indicated conditions; P values calculated with regard to mock. n = 3, minimum of 500 cells per experiment. (D) Immunoblots depicting the transient expression of the various rLIC1 phosphorylation constructs in HeLa cells. (E) Representative epifluorescence micrograph of HeLa cells stained with the viable DNA dye Syto 13 to visualize metaphase cells. Zoomed inset shows one magnified metaphase cell. Scale bar = 50 µm, inset scale bar = 10 µm. (F) Metaphase index (fraction of metaphase cells) from HeLa cells treated with anti-hLIC1 siRNA and rescued by transient expression of the various rLIC1 constructs as indicated. n = 3, minimum 600 cells per experiment. IB, immunoblot. Error bars = mean ± SEM. **, P < 0.01; ***, P < 0.001; ****, P < 0.0001, ns = not significant vs. mock (one-way ANOVA).