Extended Data Fig. 3: Regulation of myosin localization and phosphorylation in cells with nucleator or NPF depletion.
From: SPIN90 associates with mDia1 and the Arp2/3 complex to regulate cortical actin organization
a, Representative pMLC distribution in metaphase HeLa cells visualized by immuno-staining for different protein depletions and for non-silencing (NS) siRNA. Each image is a single section of a confocal microscopy stack and is shown in inverted contrast. b, Mean cortical pMLC fluorescence intensity for different treatments normalized to the mean cortical pMLC fluorescence intensity for non-silencing (NS) siRNA or shRNA. The distributions’ medians, first and third quartiles and ranges are represented by the central red bars, bounding boxes and whiskers, respectively. Statistics are derived from the total number (n, indicated above each box) of cells examined in three independent experiments. Each dot represents one cell measurement. Statistical outliers are indicated by red dots. Statistical comparisons of the means were performed using one-way ANOVA on ranks compared to NS siRNA/shRNA. ACTR2 siRNA: p = 7 10-5, NAP1: p = 0.03, mDia1: p = 0.85, IQGAP1: p = 0.08, SPIN90: p = 0.74. **p < 0.01 compared to the appropriate control. See Supplementary Figure 9 for controls. c, Change in cortical myosin regulatory light chain fluorescence intensity upon treatment with DMSO (left panel) or CK666 (right panel). In each panel, the top row shows the fluorescence intensity before treatment and the bottom row after treatment for the same cell. The left most column shows myosin regulatory light chain fluorescence (MRLC-GFP), the middle column shows LifeAct-Ruby, and the right column shows the overlay with MRLC in green and LifeAct in Magenta. Experiments were repeated twice independently with similar results. (a,c) Scale bars=10 µm. Statistical source data can be found at Source data figure ED3.
