Fig. 2: Pak3 depletion induces aberrant actin dynamics. | Nature Communications

Fig. 2: Pak3 depletion induces aberrant actin dynamics.

From: Inhibition of negative feedback for persistent epithelial cell–cell junction contraction by p21-activated kinase 3

Fig. 2

a Representative images of actin labeled with Lifeact::GFP in the A8a cells from >3 independent experiments. Arrowheads indicate some aberrant actin protrusions. Scale bar, 10 μm. b Magnified time-lapse images of Lifeact::GFP at cell–cell junctions, representative of >3 independent experiments. Scale bar, 3 μm. c Mean ± S.D. of the height and width of small actin protrusions in the control cells and aberrant protrusions in the Pak3-depleted cells. n = 26 (control and Pak3 RNAi) or 36 (Pakd02472) protrusions from 5 (Control and Pak3 RNAi) or 2 (Pak3d02472) biologically independent pupae per genotype. P-values by two-tailed Dunnett’s test. d Representative images of actin labeled with UtrABD::GFP in the A8a cells from two independent experiments. Arrowheads indicate some aberrant actin protrusions. Scale bar, 10 μm. e A schematic illustration of the large actin protrusions (magenta) at a junction (black). f Percentages of junctions generating the large actin protrusions for the indicated number of times per hour are shown. Parentheses, the number of examined junctions from 4 to 5 pupae per genotype. P-values by two-tailed Mann–Whitney U-test. g Graph showing the length of a representative junction that repeats shortening and re-extension in Pak3 RNAi cells. The timings when the large actin protrusions emerge are shown in magenta. h Mean ± S.D. of the timing of the onset of the large actin protrusion formation. Parenthesis indicates the number of protrusions from 15 junctions of 3 pupae. Genotypes: a–c +/Y;UAS-Lifeact::GFP/+;AbdB-Gal4/+ (Control) and +/Y;UAS-Lifeact::GFP/+;AbdB-Gal4/UAS-Pak3 RNAi; d +/Y;sqh-UtrABD::GFP/+ (Control) and +/Y;sqh-UtrABD::GFP/+;Pak3d02472; f +/Y;UAS-Lifeact::GFP/+;AbdB-Gal4/+ (Control), +/Y;UAS-Lifeact::GFP/+;AbdB-Gal4, UAS-H2B::ECFP/UAS-Pak3 RNAi, +/Y;UAS-Lifeact::GFP/+;AbdB-Gal4, UAS-Pak3::GFP/UAS-Pak3 RNAi, +/Y;UAS-Lifeact::GFP/UAS-Abi RNAi;AbdB-Gal4/UAS-Pak3 RNAi, +/Y;UAS-Lifeact::GFP/UAS-SCAR RNAi;AbdB-Gal4/UAS-Pak3 RNAi, +/Y;UAS-Lifeact::GFP/UAS-MRLC RNAi;AbdB-Gal4/UAS-Pak3 RNAi, and +/Y;UAS-Lifeact::GFP/UAS-Rock RNAi;AbdB-Gal4/UAS-Pak3 RNAi; g, h +/Y;UAS-Lifeact::GFP/+;AbdB-Gal4/UAS-Pak3 RNAi. Source data are provided as a Source data file.

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