Fig. 3: Pak3 depletion causes the mislocalization of myosin II cables.
a Representative time-lapse images of MRLC::GFP at the shortening junctions from >3 independent experiments. Orange and blue arrowheads indicate splitting myosin II cables and the positions of the edges of the junctions, respectively. Scale bar, 3 μm. b Graph showing the length of a junction, and the timings at which the two distinct MRLC::GFP cables split along the junctions are shown in orange. c Mean ± S.D. of timing of the onset of MRLC::GFP cable splitting along the junctions. Parentheses, the number of the MRLC::GFP cable splitting from 21 junctions of 3 pupae. d, Mean ± S.D. of the lengths of junctions when the MRCL::GFP cables initiated splitting; n = 30 junctions from 3 (Control) and 4 (Pak3 RNAi) biologically independent pupae were examined. P-value by unpaired t-test. e Representative images of cell–cell junctions with split MRCL::GFP cables (orange arrowheads), the large actin protrusions (yellow arrowheads), and both in Pak3 RNAi cells (from the left). Scale bar, 5 μm. f, g Mean ± S.D. of the percentages of the dynamics categorized in (e) at cell–cell junctions larger than (f) and smaller than (g) 3 μm. Parentheses, the number of examined junctions from n = 3 biologically independent pupae. h Representative time-lapse images of MRLC::GFP and Lifeact::Ruby at the cell–cell junction in Pak3 RNAi cells. Orange and yellow arrowheads indicate split MRLC::GFP cables and the large actin protrusions, respectively. Scale bar, 3 μm. Genotypes: a, d MRLCAX3/Y;MRLC-MRLC::GFP;AbdB-Gal4, UAS-H2B::ECFP/+ (Control) and MRLCAX3/Y;MRLC-MRLC::GFP;AbdB-Gal4, UAS-H2B::ECFP/UAS-Pak3 RNAi; b, c MRLCAX3/Y;MRLC-MRLC::GFP;AbdB-Gal4, UAS-H2B::ECFP/UAS-Pak3 RNAi; e–h MRLCAX3/Y;MRLC-MRLC::GFP/UAS-Lifeact::Ruby;AbdB-Gal4, UAS-H2B::ECFP/UAS-Pak3 RNAi. Source data are provided as a Source data file.
