Figure 4. Co-immunoprecipitation of Pax6 and Pax6-variant. A: Rabbit anti-Gfp-Pax6 antibody was used to do the IP experiment, and rabbit anti-Gfp-Pax6 antibody was used to do the western blotting. The inputs from left to right are negative control, 6-day embryonic tissue extract, Co-IP, and Pax6-his protein. Pax6 protein is obvious in the 6-day embryonic tissue extract and the Co-IP of the input, and lower than the heavy chain of the antibody. When using the rabbit anti-Gfp-Pax6-varian antibody to do the western blotting, the input from left to right is negative control, 6-day embryonic tissue extract, Co-IP, and Pax6-varian-his protein. Pax6-varian can be found in the 6-day embryonic tissue extract and the Co-IP of the input. B: The anti-Gfp-Pax6-varian antibody was used to do the IP experiment. When using anti-Gfp-Pax6-varian antibody and corresponding input, Pax6 variant was probed in the 6-day embryonic tissue extract and the Co-IP of the input; also, Pax6 was probed with an anti-Gfp-Pax6 antibody in the 6-day embryonic tissue extract and the Co-IP of the input. The results show that both Pax6 and Pax6 variant protein can be found in embryonic tissues of the Bufo raddei Strauch, and a complex of these proteins was found in the Co-IP of the input. WB: Western Blotting; WT: Wild Type.