Xenbase Image ID: 155916

Fig. 3. Mid1 induces ubiquitination and reduces the abundance of Pax6 protein. (A) Blots show levels of Pax6, Mid1, and β-tubulin in HEK293 cells after cotransfection on pax6-flag expression plasmid and either EV or myc-mid1 at different time points (min) on cycloheximide block of protein synthesis. Diagram shows the quantified values of Pax6 protein relative to β-tubulin levels in Western blot. Amounts of Pax6 protein before the cycloheximide treatment are designated as 1 (100%). (B) In vivo ubiquitination of Pax6 was analyzed by transfection of HEK293 cells with plasmids coding for pax6-flag, myc-mid1, and his-ubiquitin as indicated and anti-his Western blot of proteins eluted after an anti-flag-IP (Left). Input of the proteins was verified by Western blot of the corresponding lysates with anti-Myc and anti-Pax6 antibodies, respectively (Right). Image published in: Pfirrmann T et al. (2016) Copyright © 2016. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License. Larger Image Printer Friendly View

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