Ectopic overexpression of H19 prevents an increase in the numbers of Paneth and goblet cells in intestinal organoids isolated from H19-/-mice. (A) Levels of H19 in intestinal organoids 48 hours after transfection with an H19 expression vector or a control empty vector. Values are the means ± SEM (n = 3). *P < .05 compared with control vector. (B) Immunostaining of Paneth cells (lysozyme-positive cells, red) in intestinal organoids described in panel A: a) control; and b) H19 O/E. Scale bars: 50 μm. (C) Immunostaining of Goblet cells (mucin 2–positive cells, red) in intestinal organoids described in panel A. Scale bars: 50 μm. (D) Quantitative data of Paneth (top) and goblet cells (bottom) in the intestinal organoids described in panels B and C: a) control; and b) H19 O/E. Values are the means ± SEM (n = 12). *P < .05 compared with control vector. DAPI, 4′,6-diamidino-2-phenylindole; E-cad, E-cadherin; Lys, lysozyme; O/E, overexpression.