Figure 5
From: The Light Intermediate Chain 2 Subpopulation of Dynein Regulates Mitotic Spindle Orientation
LIC2-dynein interacts with cortical orientation complexes. (A) Still images from time-lapse movies of HeLa cell line expressing multifunctional GFP-tagged IC74 (mfGFP-IC74). Arrows indicate cortical dynein. (B) Quantification of cortical intensity of mfGFP-IC74 upon respective siRNA treatments. Y-axis represents average fluorescence intensity at the cortex, each bar represents one cell. Horizontal dashed line marks the average fluorescence intensity seen in LIC2-depleted cells. N = 26 cells each for all conditions. LIC2-depleted cells show significantly reduced mfGFP-IC74 cortical intensity in comparison to control or LIC1-depleted cells. (C) Quantification of mfGFP-IC74 intensity at the cortex upon respective siRNA treatment. Bars represent mean +/− SEM from 3 independent experiments. N = 26 live metaphase cells from 3 independent experiments). (D) Affinity purification of SBP-tagged empty vector (control), LIC1 and LIC2 followed by immunoblotting with respective antibodies as indicated. IN = input, FT = flow through, WS = wash, AP = affinity purified eluate.
