Ad-siHOPX silences HOPX in culture and leads to increased size of spheroids grown from 3-h ischemic-injured (3hrI) crypts. A: HOPX expression is significantly decreased in control crypts transduced with Ad-siHOPX compared with Ad-Sc at 48 h in culture. Fold changes were calculated relative to Ad-Sc samples and GAPDH as housekeeping gene. Statistics analyzed by two-way ANOVA with *P = 0.05, n = 8. B: representative images of spheroids at 24, 48, and 72 h in culture, grown from no virus, Ad-Sc, and Ad-siHOPX transduced 3hrI crypts. Transduction confirmed by GFP expression (green). Scale bar 20 µm. C: box plots of log transformed daily spheroid area measurements from no virus (green), Ad-Sc (blue), and Ad-siHOPX (red) transduced 3hrI and control crypts, *P < 0.05 **P < 0.01 ***P < 0.001, n = 5 pigs, 15–20 spheroid/enteroids per condition measured.