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. Author manuscript; available in PMC: 2021 Oct 15.
Published in final edited form as: Cell. 2020 Oct 1;183(2):395–410.e19. doi: 10.1016/j.cell.2020.08.045

Figure 4. Epigen Is Stored and Concentrated within Intercellular Nanolumina.

Figure 4.

(A) TEM images of MMTV-PyMT clusters (pseudo-colored). MP, microvilli-like protrusions; J, cell-cell junction.

(B) Models of intercellular epigen signaling as a membrane-restricted juxtacrine ligand or a diffusible paracrine ligand.

(C) Epigen immunofluorescence stimulated emission depletion microscopy images in a MMTV-PyMT cluster.

(D) Immunogold labeling of epigen in MMTV-PyMT clusters. Arrows, gold labeling.

(E) Schematic of features scored for Immunogold signal. Right: quantification of epigen Immunogold signal. n = 268 gold particles.

(F) To assess junctional permeability, clusters were incubated with sulfo-NHS-biotin. EGTA was used as a positive control to disrupt cell-cell junctions. Right: quantification of streptavidin-fluorescein isothiocyanate (FITC) fluorescence. n = 3 biological replicates. n = 75 clusters per condition. p values = Mann-Whitney tests.

(G) Immunofluorescence of epigen in clusters ± 1 mM EGTA treatment. Right: median epigen intensity per cluster. n = 3 biological replicates, 59 clusters. p value = Mann-Whitney test.

See also Figures S4 and S5.