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. 2013 Jul 21;4(8):1266–1279. doi: 10.18632/oncotarget.1164

Figure 6. Effects of PTEN on PriGO8A differentiation.

Figure 6

A. PriGO8A cells were transduced with Tet activator lentivirus and selected with G418 for four days. Selected cells were then transduced with inducible PTEN lentivirus and selected with puromycin for four days. Selected cells were treated with the indicated concentrations of doxycycline for 48 h. Total cell lysates were then analyzed by Western blotting for expression of PTEN. B. PriGO8A cells with inducible PTEN were treated with 500 ng/ml doxycycline for two days and then either mock transduced or transduced with lentivirus constitutively expressing wild-type Lgl1. One day later cells were switched to media without EGF. Total cell extracts were collected one day later and analyzed by Western blotting with antibodies to phospho-PKC substrate and Flag epitope. C. PriGO8A cells with inducible PTEN were untreated or treated with 500 ng/ml doxycycline for 7 days. Cells were then fixed and immunocytochemistry for TUJ1 and GFAP expression was performed. D. Quantitation of data from C. The left graph shows doxycycline treatment of PriGO8A cells with inducible PTEN. The right graph shows doxycycline treatment of parental PriGO8A cells (as a control for effects of doxycycline alone). Quantitation was performed as described in Material and Methods. Data are shown as the mean ± SE. * indicates a p value less than 0.05.