Figure 5
S12 is essential for cytokinesis.
(a) HeLa cells were stably transfected with HA vector, HA-ATXN10-res and HA-ATXN10-12A-res, S77AT82A (2A) -res, S12AS77AT82A(3A) -res plasmids and then treated with control siRNA or siATXN10, as indicated. The lysates were collected 48 hrs later and subject to IB using the antibodies indicated. (b) the same cells in (a) were analyzed by immunofluorescence for multinucleated cells and the data were quantified. The data are representative of two independent results. More than 100 cells were counted in each sample and asterisks indicate significant differences as determined by χ2 test. (d) HeLa cells ectopically expressing HA-Ataxin-10, S12A, 2A, or 3A mutants were costained with anti-HA and anti-α-tubulin antibodies. Binucleated or multinucleated cells were scored. Scale bar, 10 μm. (e) The histogram shows the mean ± SD from three independent experiments. More than 100 HA-positive cells were counted in each experiment. Asterisks indicate significant differences from wild type (p < 0.05); double dots indicate significant differences from S12A (p < 0.05), as determined by t-test (p1–2 = 0.007, p1–3 = 0.039, p1–4 = 0.00017, p2–3 = 0.129, p2–4 = 0.008, p3–4 = 0.39). Uncropped images of blots were shown in supplemental Figure S1.
