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Figure 6


Fig. 6. Functional expression of {alpha}3 and {alpha}8-G22R connexins in Xenopus oocytes. (A) Western blot analysis of Xenopus oocytes shows similar levels of {alpha}8-G22R and {alpha}3 proteins when expressed alone or co-injected. Equal amounts of oocyte membrane fractions were loaded into each lane, and the blots were probed with a polyclonal anti-{alpha}8 connexin (left), or anti-{alpha}3 connexin (right) antibody. Similar levels of {alpha}8-G22R and {alpha}3 protein synthesis were seen in individually and co-injected oocytes. Quantitation of band intensity by densitometry confirmed that levels of connexin expression were not statistically different (P>0.05). Densitometry values are the mean of three independent experiments. (B) {alpha}8-G22R connexin forms functional heteromeric channels with {alpha}3 connexin. Junctional conductance values were measured between oocyte pairs injected with wild-type {alpha}3, {alpha}8-G22R, co-injected {alpha}8-G22R and {alpha}3, or water. Oocyte pairs containing wild-type {alpha}3 alone (n=20), or co-injected {alpha}8-G22R and {alpha}3 (n=57) exhibited conductance values >100-fold higher than the pairs injected with water for the background value (n=40). However, conductance of co-injected {alpha}3 and {alpha}8-G22R was reduced ~75% compared with {alpha}3 alone (P<0.05). Homotypic {alpha}8-G22R (n=55) cell pairs failed to produce levels of coupling that were significantly higher than background (P>0.05). Similarly, heterotypic {alpha}8-G22R/{alpha}3 cell pairs, formed by pairing one oocyte expressing {alpha}3 with another injected with {alpha}8-G22R, were poorly coupled (n=16).





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