Fig. 3: IL-8/G-CSF secreted by Scar transformed decidual fibroblasts chemotactically recruit EVTs.
From: Scar matrix drives Piezo1 mediated stromal inflammation leading to placenta accreta spectrum
A Heatmap showing significant differential ligand encoding genes expressed in dESFs on Scar and Physio matrices. B Representative IL-8 immunofluorescence images of dESFs treated with protein transport inhibitor GolgiStop for 6 h on Physio and Scar matrices, quantification shown in right panel; n = 49 and 53 cells for Physio and Scar, respectively. p = 5 × 10−11. C, D ELISA based analysis of IL-8 and G-CSF concentration in supernatant of dESFs on Physio and Scar in 3D and 2D; n = 3 samples. p = 0.0007 in (C) and p = 0.04 and 2 × 10−5 in (D). E ELISA based measurement of IL-8 concentration in supernatant of dESFs treated overnight with DMSO, or 100 ng/ml TNFα; n = 3 samples. p = 0.0097. F Experimental workflow to test migration of HTR8 in medium conditioned from dESFs with gene silenced for IL-8 and G-CSF encoding genes, CXCL8 and CSF3, respectively. G Migration trajectories (initial location (x, y = 0,0)) of HTR8 conditioned with medium from dESFs silenced for CXCL8 and CSF3 genes, without or with addition of recombinant human (rh) IL-8 and G-CSF; Quantification of averaged velocities over 24 h shown in (H); p = 1×10−8, 8×10−7, 8×10−9, and 6 × 10−5; n listed below each condition. I 3D chemotaxis of primary EVTs in collagen gel towards IL-8 and G-CSF gradient; Shown is a representative image of EVTs in collagen gel (left); Trajectories of individually tracked EVTs from their initial location (0,0) (middle and right); Cell trajectory with mean displacement towards cytokine end are labeled red, and counted (n); p value showing Rayleigh test of cell trajectories: p < 0.05 is considered chemotaxis. J ANSIA-based stromal invasion analysis of HTR8 in monolayer of dESFs silenced for CXCL8 and CSF3 genes; Control refers to scrambled sgRNA. p = 0.003 and 0.005. Data in figures B–E, H and J are showing as mean ± s.d.; statistical significance is determined by unpaired two-tailed t-test (*p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001). Source data are provided as a Source Data file. Figure 3F created with BioRender.com released under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International license (https://creativecommons.org/licenses/by-nc-nd/4.0/deed.en).
