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Figure 8


Fig. 8. Affinity purification of HA-tagged connexin and potentially associated proteins. The presence and abundance of Cx40, Cx43 or Cx26 were detected in the cell extract, washes and eluted material by immunoblotting using anti-connexin antibodies. Gels were loaded with 25 µg of protein (corresponding to 1/10 of the amount incubated with MicroBeads) in the lane labeled `Before column'; 1/10 of the fraction collected as a `Flow through', 1/5 of each of the `Washes'; and 1/10 of the `Eluate' (corresponding to 25 µg of protein applied to the purification column). (A) Purification of proteins from HeLa-Cx43 cells transfected with Cx40HA. (B) Purification of proteins from HeLa-Cx43D12S,K13G [Cx43 (mut)] cells transfected with Cx40HA. (C) Purification of proteins from HeLa-Cx26HA cells transfected with Cx43. (D) Purification of proteins from HeLa-Cx26HA cells transfected with Cx43D12S,K13G. (E) Purification of proteins from HeLa-Cx40 cells transfected with Cx26HA. (F) Purification of proteins from HeLa-Cx40E12S,E13G [Cx40 (mut)] cells transfected with Cx26HA.