Fig. 5. Inhibiting RhoA signaling promotes SCN axon turning to netrin 1. (A,B) Aggregates of netrin 1-expressing HEK 293-EBNA cells were cultured alongside explants of E11 rat spinal cord and deflection distances measured as shown (A), and graphed (B) as the percent distance relative to the absence of drugs. (C-H) Explants were cultured for 40 hours in the absence of RhoA inhibitors (C,D), or in the presence of 10 µg/ml C3-07 (E,F) or 10 µM Y-27632 (G,H). SCN axons were fluorescently labeled for Tag1 (4D7). Both C3-07 (E, n=30, P=0.005) and Y27632 (G, n=30, P=0.007) resulted in a 16% increase in the turning distance. DCC function blocking antibodies (5 µg/ml) reduced turning in the absence of drug (D, n=24, P=0.001), as well as turning in the presence of 10 µg/ml C3-07 (F, n=21, P<0.001) or 10 µM Y27632 (H, n=24, P<0.001). Tukey post-hoc tests of means. Error bars indicate s.e.m. ^**P<0.001 compared with control explants without Y27632, C3-07 or DCC-fb. Scale bar: 20 µm.