Fig. 4. Increased clone size upon overexpression of mouse Par3 in vitro. (A-D) GFP-labeled clone in E12 cortex dissociated-cell cultures after Par3 transduction and 7-days culture. Immunolabeled for GFP (A and D, green), nestin (C and D, blue) and MAP2 (B and D, red). Note the high number of nestin-positive MAP2-negative cells. (E-G) Bar charts representing the size (E,F) and neuronal content (G) of clones generated from E12 cortical cells after transduction with control or Par3-containing retroviral vectors. Note the significant increase in the average clone size (F; ^***, P<0.001; unpaired t-test), as well as in the frequency of clones containing 20 or more cells (E; ^*, P<0.05; unpaired t-test) after Par3 transduction (n=132) in comparison to the control (n=170). The proportion of neurons amongst GFP+ transduced cells was also significantly reduced after Par3 gain-of-function (G; ^**, P<0.01; unpaired t-test). Scale bar: 50 µm.