Fig 2. Co-IP experiment confirms that Cx43 interacts with F-actin through drebrin binding.
(A) Schematic of the co-IP strategy: F-actin antibody was bound on protein A agarose beads to capture F-actin, serving as the “bait” protein. GST-Cx43-CT240-382 and GST-Drb1-300 purified constructs were used as “prey” proteins. (B) Representative lanes of Western blots chosen as best example from four independent experiments performed with a rabbit anti-GST antibody. The arrows indicate the GST-Cx43-CT240-382 (blue) and GST-Drb1-300 (green) bands obtained with or without the addition of F-actin (lanes 2, 4, and 5, and 1 and 3 respectively). (C) Quantification of the data shown in (B). GST-Cx43CT240-382 or GST-Drb1-300 co-IP band intensities were normalized to the band obtained without the “bait” protein (F-actin). The p value is the result of the T-test ran to confirm significant difference between the two samples. Error bars represent the standard deviation between four replicates.