Fig. 6.

The role of P2X7 receptors in mouse ES cell colony number and self-renewal. Mouse ES cells were plated at approximately 166 cells/cm² and cultured for 5 days (in the presence or absence of 10 μM A438079) prior to counting the percentage of colonies intensely staining for pure alkaline phosphatase activity (A–B) and total colony number normalised to control untreated cells (C). Cells cultured in GMEM or KO-DMEM (indicated in the figure) for 5 days as described in Materials and methods prior to analysis for colony number (C) and frequency of pure alkaline phosphatase positive colonies (B) where data are plotted as the mean ± SEM (n = 3). (A) A representative image of mES cells cultured in KO-DMEM in the presence or absence of 10 μM A438079. Scale bar = 400 μm. (D) Nanog-GFP expression in mES cells cultured in KO-DMEM for 48 and 72 h in the absence or presence of 10 μM A438079 as indicated in the figure. GFP fluorescence analysed by flow cytometry, representative images in figure with GFP^-ve control cells in insert. (E) Mean GFP fluorescence from flow cytometry experiments plotted as a histogram with n values indicated in the figure. Data plotted as mean ± SEM.