Skip to main content
. 2016 Mar 4;44(11):5105–5122. doi: 10.1093/nar/gkw113

Figure 1.

Figure 1.

Combination of NGF and cAMP stimulations caused cell cycle arrest in differentiated PC12 cells. (A) Morphologies of PC12 cells that grew during 7 days of differentiation in the presence of NGF (upper) or NGF/cAMP (lower). (B) Relative number of living cells after removal of NGF or of NGF and cAMP. Upper diagram shows experimental scheme. After 7 days of differentiation of PC12 cells, NGF and cAMP were removed to allow proliferation. Number of living cells was evaluated by counting trypan blue staining-negative cells. Number of cells in each differentiation state at day 7 was set as 1. (C) Cell proliferation monitored by EdU incorporation and Ki67 staining in undifferentiated (Undiff), NGF-differentiated (Ndiff) and NGF/cAMP-differentiated (NcAdiff) cells, and in Ndiff and NcAdiff cells after 4 days of incubation in proliferative condition (Ndiff + reverse 4 day, NcAdiff + reverse 4 day, respectively). (D) Proportion of apoptotic cells before/after removal of NGF or NGF and cAMP, detected by active Caspase 3 (aCas3) staining. In (B–D), values are mean ± SD (n = 3). Statistical analysis was performed using Student's two-tailed t-tests, or the Tukey-Kramer multiple comparison test. N.S. = not significant. *P < 0.05; **P < 0.01; ***P < 0.001.