Fig. 6

Inhibition of Rho kinase, JNK, and cytoskeletal architecture negatively affects biliary development. (A–F) Whole-mount projections of cytokeratin immunostaining of liver from a 5 dpf control larvae (A), compared to similar stainings from larvae treated with the Rho kinase inhibitors fasudil (B) and H-1152 (RKI, C), as well as the JNK inhibitor dicoumeral (D), the actin inhibitor cytochalasin D (E), and the microtubule and cytoskeleton inhibitor colchicine (F). Note that inhibition of any of the above downstream targets of PCP leads to a phenotype similar to the pk1a morphant phenotype. (G) Bar graph depicting PED6 treatment of control larvae compared to larvae treated with low dose pk1a morpholino, low dose cytochalasin D, or the combination. Note that alone, the low dose MO or cytochalasin have no to minimal effect, but that in combination, the effect is sizable and significant (p < 0.0005, chi-square). (H) Bar graph depicting PED6 treatment of control larvae compared to larvae treated with low dose pk1a morpholino, low dose colchicine, or the combination. Note that only in the combination is there a statistically significant effect on PED6 gallbladder uptake (p < 0.0001, chi-square test). (I–N) Confocal projections of cytokeratin immunostaining of livers from 5 dpf control larvae (I) and larvae injected with low dose (l.d.) pk1a MO (J), low dose cytochalasin D (cyto D, K), low dose colchicine (colch, L), and low dose pk1a MO with low dose cytochalasin D (M) or low dose colchicine (N). Note that (J–L) appear similar to control (I), but that (M, N) are abnormal.