Fig. 1: PTEN loss enhances collective cell migration independently of PI3K/AKT signalling. | Nature Communications

Fig. 1: PTEN loss enhances collective cell migration independently of PI3K/AKT signalling.

From: PTEN inhibits AMPK to control collective migration

Fig. 1

a Phase-contrast images of siCTL and PTEN-depleted (siPTEN#1,2) astrocytes migrating collectively in a wound-healing assay. Scale bar: 100 μm. b Diagrams showing 24 h-trajectories of 10 representative siCTL, and siPTEN#1 cells rescued with GFP alone (Ø), PTEN-WT(WT) and PTEN-C124S (C124S). c Mean astrocyte velocity. 69 siCTL cells, 107 siPTEN + Ø cells, 133 siPTEN+wt cells, 133 siPTEN+C124S cells, 145 siPTEN+G129E cells and 137 siPTEN + Y138L cells from three biologically independent experiments were examined. Statistics were drawn using two-tailed Mann–Whitney test. p values and effect sizes vs siCTL are given in the table above the graph. d Schemes representing lateral and dorsal view of a 24/30hpf zebrafish embryo with dorsal (DA) and caudal arteries (CA) in red and the caudal (CV) and common cardinal (CCV) veins. CCV endothelial cells (ECs) are delaminating from the midline, moving towards the heart (yellow box). e Representative fluorescent image of migrating ECs expressing Lifeact-eGFP. Scale bar: 50 μm. The micrograph is representative of at least 12 independent fish larvae. f Time-colored zoomed-in image of migrating ECs. Red is t = 0, blue is t = 20 min, yellow is t = 40 min and white is t = 180 min. Scale bar represents 10 µm. g, h Mean velocity of lifeact-eGFP expressing ECs in control zebrafish embryos (MoCTL), ptena morphant (MoPTENa) and ptenb morphant (MoPTENb) (g, n = 18, 17, 13 cells; N = 4 fish, two-tailed unpaired t-test) and in wild-type pten and in ptenb−/− mutant zebrafish embryos (h, n = 19, 21 cells; N = 6, 5 fish; two-tailed unpaired t-test). i Phase-contrast images of siPTEN#1 cells treated with DMSO or the PI3K inhibitor LY294002. White dashed lines delineate the border of the wound at t = 0. Red lines delineate the border of the monolayer 24 h later. Scale bar 100 μm. j, k Mean velocity of siCTL and siPTEN cells treated with or without LY294002 (j, n = 198 cells, N = 3, two-tailed Mann–Whitney test) and with DMSO or VO-OHpic (k, n = 300, N = 3, two-tailed Mann–Whitney). Error bars represent Standard Deviation (SD). Boxes for box-plot graphs (g, h) extend from the 25th to 75th percentiles and the line in the middle is plotted at the median. Whiskers delineate all data points from minimum to maximum. Source data are provided as a Source Data file.

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