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. 2014 Dec 10;9(12):e114964. doi: 10.1371/journal.pone.0114964

Figure 3. High-molecular weight p62 induced by singlet oxygen production.

Figure 3

(A) 50 ng GST-p62 was incubated with 0.05% DMSO, 5 µM verteporfin, or 5 µM rose bengal for 1 h at 4°C or 37°C in the dark. (B) Immunoprecipitated p62 from untreated BxPC-3 cells was exposed to 0.1% DMSO, 10 µM verteporfin, or 10 µM rose bengal for 30 min in the dark at 4°C or 37°C or to (C) 0.1% DMSO, 10 µM verteporfin, or 10 µM rose bengal in the dark at 37°C with or without 20 mM histidine. Reactions were immunoblotted for p62. (D) 26 ng purified His-p62 was incubated with 0.05% DMSO, 5 µM verteporfin, or 5 µM rose bengal for 1 h at 4°C or 37°C in the dark or (E) 50 ng purified GST-p62 was incubated with 0.1% DMSO, 10 µM verteporfin, or 10 µM rose bengal for 30 min at 37°C in the dark. Samples were derivatized with DNPH, subjected to SDS-PAGE, and immunoblotted with anti-DNP antibody solution according to the Oxyblot kit manufacturer’s instructions. All images presented are representative of at least 3 independent experiments.