Fig. 1: Pak3 is required for cell–cell junction shortening during epithelial junction remodeling. | Nature Communications

Fig. 1: Pak3 is required for cell–cell junction shortening during epithelial junction remodeling.

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

Fig. 1

a Top, representative time-lapse images of male genitalia observed from the posterior end during rotation from >3 independent experiments. The ventral side is located at the top. Nuclei in the A8a cells and the whole body are depicted in green and purple, respectively. Yellow and white arrows indicate the movements of genitalia and the A8a epithelia, respectively. Scale bar, 100 μm. Bottom, a schematic illustration showing the angles of genitalia with the A8p epithelia (both purple) and the A8a epithelia (green), and a schematic illustration of cell intercalation in the A8a epithelia. Orange lines indicate shortening junctions. b Top, a schematic illustration of the categorization of genitalia angles. Yellow arrow indicates genitalia movement. Gray indicates the 360° rotation of genitalia (normal orientation). Bottom, percentages of male adult flies with the indicated genitalia angles are shown. Parentheses, the number of males examined. c Representative images of A8a epithelia at 28 h APF immunostained for Pak3 and Dlg from two independent experiments. Pak3 RNAi cells are indicated with mCherry signals. Scale bar, 10 μm. Broken lines indicate the edges of Pak3 RNAi clones. d Representative time-lapse images of E-Cad::GFP at remodeling junctions from >3 independent experiments. Stars and triangles indicate cells forming the shortening junctions. Orange, blue, and green arrowheads indicate shortening junctions, four-way vertices, and growing junctions, respectively. Scale bar, 5 μm. e A schematic illustration of the categorization of the junction dynamics. f Mean ± S.D. of the percentages of junctions with the categorized dynamics. Bar colors correspond to e. Parentheses, the number of examined junctions from n = 3 biologically independent pupae per genotype. P-values by two-tailed Dunnett’s test. Genotypes: a +/Y;His2Av::mRFP/+;AbdB-Gal4, UAS-H2B::ECFP/+ (Control) and +/Y;His2Av::mRFP/+;AbdB-Gal4, UAS-H2B::ECFP/UAS-Pak3 RNAi; b (bottom left) +/Y;;AbdB-Gal4/+ (Control), +/Y;;AbdB-Gal4/UAS-MRLC RNAi, and +/Y;;AbdB-Gal4/UAS-Pak3 RNAi; (bottom right) +/Y;UAS-Lifeact::Ruby/+;AbdB-Gal4/UAS-Pak3 RNAi (Control), +/Y;UAS-Dicer2/+;AbdB-Gal4/UAS-Pak3 RNAi, and +/Y;;AbdB-Gal4, UAS-Pak3::GFP/UAS-Pak3 RNAi; c hs-flp/Y;E-Cad::GFP (KI);Act > CD2 > Gal4, UAS-mCD8::mCherry/UAS-Pak3 RNAi; d, f +/Y;E-Cad::GFP (KI);AbdB-Gal4, UAS-H2B::ECFP/+ (Control), +/Y;E-Cad::GFP (KI);AbdB-Gal4, UAS-H2B::ECFP/UAS-Pak3 RNAi, and +/Y;E-Cad::GFP;Pak3d02472. Source data are provided as a Source data file.

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