Fig. 6.

Increasing hLRH-1 in human intestinal organoids protects against TNFα. a Brightfield view of human small intestinal organoid. Scale bar = 100 μm. b Immunofluorescence for LRH-1 (green, top panels) in human intestinal organoid sections shows expression throughout the organoid with strongest expression occurring in the crypt domain (yellow dashed box and zoomed image, right). Differentiation markers for Paneth (Lyz, left) and goblet (Muc2, right) cells are shown below. Scale bar = 100 μm. c Expression of LRH-1 target gene Ctrb1 in human intestinal organoids is upregulated 72 h after infection with AAV-hLRH1 (blue) but not with control AAV (black). d Overexpression of hLRH-1 by AAV confers resistance to TNFα-mediated cell death. Human organoids from healthy donor and a Crohn disease patient were infected with AAV-hLRH (blue) or AAV-Control (black) (3.3×10¹⁰ genome copies) for 72 h under differentiation conditions and then exposed to TNFα (20 ng/ml, 40 h). Data represent an N of at least three replicates with ~50 organoids per well. For panels c and d error bars are SEM with statistical analyses determined by Student’s unpaired t test, two tailed with p values of *p = < 0.05