Figure 4: The dynamic perinuclear actin network is nucleated by Arp2/3. | Nature Communications

Figure 4: The dynamic perinuclear actin network is nucleated by Arp2/3.

From: Perinuclear Arp2/3-driven actin polymerization enables nuclear deformation to facilitate cell migration through complex environments

Figure 4

(a) Percentage of cells with F-actin accumulation in 2-μm-wide constrictions after nuclear entry. n, no; y, yes. Numbers represent the number of cells scored. ***P<0.0001 by Fisher’s test. (b) Representative images of LifeAct-GFP (green)-expressing DCs stained with Hoechst (red) passing or failing to pass a 2-μm-wide constriction. (i) Time point before cell and nuclear entry in constriction; (ii) time point after cell but before nuclear entry in constriction; (iii) time point after cell and nuclear entry in constriction. Scale bar, 10 μm. (c) Left graph: normalized mean actin intensity in DCs that succeed (green) or fail (red and purple) to pass 2-μm-wide constrictions as a function of nuclear front position centred on the constriction. Grey bar represents the constriction. Right graph: zoom on constriction entry (region R1) of the left graph. (d) Normalized mean actin intensity in DMSO-treated (green) and 50 nM of Latrunculin A-treated (red) DCs as a function of nuclear front position centred on the constriction. Grey bar represents the constriction. (e) Normalized mean actin intensity in constriction measured in DMSO-treated (green) and 50 nM of Latrunculin A-treated DCs as a function of the nuclear entry and exit time. (c,d,e) Dark and light colours respectively represent the mean and the s.d. (f) Representative image of a DC immunostained for Arp2/3 and stained for actin filaments in a 2-μm-wide constriction. Scale bar, 15 μm (left); 5 μm (right). The pinched DNA at the actin accumulation site is noteworthy. (g) LifeAct-GFP (green)-expressing DCs stained with Hoechst (red) in 2-μm-wide constrictions. Left: control cell; right: after Arp2/3 inhibition. Scale bar, 15 μm. (h) Sequential images of a representative myosin IIA-GFP (cyan)-expressing DC stained with Hoechst (red). Scale bar, 15 μm (left); 5 μm (right). (i) Normalized mean myosin IIA-GFP (blue) intensity in 2-μm-wide constrictions. L, constriction length; n, number of cells; W, constriction width. N≥2. All error bars are s.e.m.

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