Fig. 1: Changes in PLCβ1 levels in NT2 cells during differentiation by RA and AraC.

A NT2 cells progenitors were treated with RA and immunoblotted with anti-PLCβ1a and PLCβ1b antibodies at various times (48 h, 7 d, 14 d, 28 d). Terminally differentiated NT2N cells were isolated from cultures at 28 d of RA exposue by mechanical dislogding (see Methods). Equal amounts of total protein (12 μg) were loaded (n = 4). B Similar studies as in (A) except NT2 progenitors were treated with AraC at various times (12, 24, 48 and 72 h and 6 days -NT2N cells-) where time points were chosen from previous studies [3]. Equal amounts of total protein (12 μg) were loaded. The optical density (OD) is expressed as the percentage of immunoreactive signal found in NT2 progenitor cells (100%). Repeated measures one-way ANOVA followed by Tukey’s post hoc test. Significant differences between the different samples with respect to NT2 progenitors are shown (*p < 0.05; **p < 0.01; ***p < 0.001). Data are mean ± SEM (n = 4). C Immunoblots of PLCβ1 on whole homogenates of NT2 progenitors, AraC-treated cells, and AraC/NT2N cells and NF200 and βIII-tubulin. Equal amounts of total protein (12 μg) were loaded.