Fig. 7.

Cellular proliferation is regulated by Cx32 acetylation. BrdU incorporation by N2a cells expressing the pCx32-IRESEGFP WT, 5R, or 5Q constructs were quantified by immunofluorescent staining for Cx32 and BrdU, as described in methods. Following overnight incubation with medium containing BrdU, cells were fixed, labeled and scored for Cx32 expression and BrdU incorporation. The percentage of BrdU-positive and Cx32 co-expressing cells were normalized to the total number of Cx32-expressing cells for each construct. a The percentage of BrdU positive cells in 5R samples were significantly reduced compared to WT (*p < 0.0026). b No differences in the percentage of Cx32 positive cells per total DAPI positive nuclei (cell number) were noted. For each construct, three to four fields were imaged and quantified for each replicate coverslip per experiment using the MultiWavelength Cell Scoring app in Metamorph, as described in the Methods. (N = 3 experiments for each construct). c Representative images of BrdU incorporation by N2a cells expressing the pCx32-IRESEGFP WT, 5R, or 5Q constructs, labeled and processed as described above