Suppression of TRTS plus SP600125-mediated neuritogenesis by U0126 in PC12-P1F1 cells. (a–d) PC12-P1F1 cells were treated with 40 ng/mL bone morphogenetic protein 4 (BMP4) as a control for 7 days in the presence or absence of U0126. Representative phase-contrast images of PC12-P1F1 cells on day 7 after treatment with (a) 0 μM, (b) 0.31 μM, (c) 0.63 μM, and (d) 1.25 μM U0126. (e–h) PC12-P1F1 cells were exposed to TRTS plus SP600125 for 7 days in the presence or absence of U0126. Representative phase-contrast images of PC12-P1F1 cells on day 7 after treatment with (e) 0 μM, (f) 0.31 μM, (g) 0.63 μM, and (h) 1.25 μM U0126. Scale bars: 100 μm. (i,j) PC12-P1F1 cells were stimulated with 40 ng/mL BMP4 (i) or exposed to TRTS plus SP600125 (j) for 7 days, and the rate of neurite-bearing cells was determined on day 7. The data represent the means ± standard deviation of three replicates. †† p < 0.01 vs. control (i) or TRTS alone (j); ** p < 0.01 vs. BMP4 alone (i) or vs. TRTS plus SP600125 only (j); n.s., not significant vs. TRTS alone (j).