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. Author manuscript; available in PMC: 2015 Aug 1.
Published in final edited form as: FEBS J. 2014 Jul 1;281(15):3357–3381. doi: 10.1111/febs.12866

Fig. 5.

Fig. 5

Sensitive Sumo-ELISA/Sandwich-ELISA assays validated that both extrinsic and intrinsic Prdx6 were Sumoylated, and showed that a fraction of endogeneous Prdx6 was present in Sumoylated form. (A) hLECs were transfected with pEGFP-vector or pEGFP-vector plus pEGFP-Sumo1 or pGFP-Prdx6 plus pEGFP-Sumo1. After 48h total cell lysates were prepared and submitted to Sandwich/Sumo1-ELISA assays to check the total Prdx6 protein and Sumoylated Prdx6 protein. Sumoylated Prdx6 protein was subtracted from total Prdx6 protein, presenting as Prdx6 unSumoylated (gray bars) and Sumoylated (black bars) forms. The data represent mean ± SD from three independent experiments (*p<0.001). (B) Prdx6+/+ LECs were transfected with pEGFP-vector plus pEGFP-Sumo1 or pGFP-Prdx6 plus pEGFP-Sumo1. Total cell lysates were prepared and used to perform Sandwich/Sumo1-ELISA. Sumoylated Prdx6 protein was subtracted from total Prdx6 protein, presenting as Prdx6 unSumoylated (gray bars) and Sumoylated (black bars) forms. The data represent the mean ± SD of three independent experiments (*p<0.001 vs control).