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. 2020 May 20;9:e54469. doi: 10.7554/eLife.54469

Figure 5. The interaction with HSPGs potentiates the ability of RSPO3 to support the growth of small intestinal organoids.

Figure 5.

(A) Bright-field microscopy images of B6 mouse small intestinal organoids grown in EN medium supplemented with various purified RSPO3 proteins at the indicated concentrations. Green arrowheads indicate viable, crypt-containing organoids and red asterisks mark dead organoids. The scale bar in the bottom right image represents 100 μm. (B) The viability of organoids grown at various concentrations of the indicated RSPO3 variants was quantified from images of the type shown in (A). Each circle represents the quantification of an independent experiment. The inset (right) shows a magnified view of organoid viability at a ligand concentration of 0.1 nM. (C) Confocal microscopy images of mouse small intestinal organoids grown in medium supplemented with 1 nM of the indicated purified RSPO3 proteins. DAPI (blue) stains nuclei, phalloidin (red) stains actin filaments at the apical surface of cells, lysozyme (green, top row) stains Paneth cells and Ki67 (green, bottom row) stains proliferating cells. For both images in each row, the scale bars in the right image represent 30 μm.