The pattern of heterotypic communication in primary astrocytes is identical to HeLa cells. Lentiviral transduction of astrocytes was at least as efficient as in HeLa cells allowing experiments to be completed well before (≤15 DIV) the appearance of endogenous Cx30 (21-28 DIV). As with HeLa cells, neurobiotin (NB) transfer was not observed in pairings of Cx47-Cx32 (A), Cx47-Cx26 (B), or Cx32-Cx43 (C) but was readily detected in pairings of Cx47-Cx43 (D), Cx47-Cx30 (E), Cx32-Cx30 (F), and Cx32-Cx26 (G). Scale bar: 50 μm.