Fig. 2: Basal actin star assembly creates a multicellular connecting network.
From: A multicellular star-shaped actin network underpins epithelial organization and connectivity
A Airyscan microscopy and depth-coded z-projection of actin, membranes and DNA localization in the basal side of organoid-derived monolayers. Scale bar, 10 μm. B N-SIM (Structured Illumination Microscopy) analysis of basal actin distribution in organoid-derived monolayers. Scale bar, 15 μm, inset 2 μm. C Statistical analysis of the distance between the AcS node and the cell centre of mass in organoid-derived monolayers. Mean distance = 1.48 ± 1.46 μm (mean ± SD). N = 3 experiments, n = 113 cells. Source data are provided as a Source Data file. (D) Statistical analysis of the number of actin branches per AcS. Mean number of actin branches per star = 6.04 ± 0.67 (mean ± SD). N = 3 experiments, n = 90 cells. Source data are provided as a Source Data file. E Statistical analysis of the actin cable length in AcSs. Mean length = 5.16 ± 1.13 (mean ± SD). N = 4 experiments, n = 331 cells. Source data are provided as a Source Data file. F Statistical analysis of the angle formed between actin cables (α) or between actin cables and plasma membranes (β). Mean α = 59 ± 15.75° (mean ± SD), mean (β) = 89.3 ± 13.13°. N = 4 experiments, n (α) = 162 cells, n (β) = 55. Source data are provided as a Source Data file. G Airyscan analysis of actin (green), membranes (magenta) and DNA (blue) in organoid-derived monolayers. Scale bar upper row 5 μm, low row 2 μm. Ha–e Transmission electron microscopy analysis of the basal domain of differentiated cells in organoid-derived monolayers. Actin cables are pseudo-coloured in green, adherens junction-like adhesions in light blue. ECM extracellular matrix. Non-pseudo-coloured images are shown in Supplementary Fig. 2C. Scale bars, a 5 μm, b, c 2 μm, d, e 0.5 μm. I and J Confocal analysis of E-cadherin and α-actinin-1 at the basal side of organoid-derived monolayers. Lower panel shows zoomed-in representative image of cell-cell junctions. Scale bar, 5 μm. K Scheme describing the architecture of AcS network (green) and cell membranes (magenta) in the basal side of organoid-derived monolayers. E-cadherin-based complexes observed in H and I are depicted in blue. α, angle formed between actin cables. β, angle formed between actin cables and plasma membranes.
