B4 knockdown affects proliferation and apoptosis. A) Decrease of B4 protein after B4 morpholino injection. After lentectomy, B4 vivo-morpholino was injected intraperitoneally every day in N. viridescens. On d 10, dorsal irises were collected, and the amount of B4 protein in the irises was measured by Western blotting. Amount of B4 was normalized with that of histone H3. *P = 0.000310 (control, n=2; B4, n=2). B–F) Effect of B4 knockdown in lens regeneration. B) Size of regenerated lens. *P = 0.00262 (control, n=7; B4, n=9). C) Number of BrdU-positive cells. *1: P = 0.0176 (control, n=4; B4, n=7); *2: P = 0.0000972 (control, n=7; B4, n=10). D) Number of Tunel-positive cells. *P = 0.00738 (control, n=6; B4, n=6). Error bars = sd. P values are from Student’s t test (2-tailed). E) BrdU immunostaining of lens from morpholino-injected newts 20 d after lentectomy. F) Tunel staining of lens from morpholino-injected newts 20 d after lentectomy. Scale bars = 100 μm. Cont. Morph., control vivo-morpholino; B4 Morph., B4 vivo-morpholino.