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. Author manuscript; available in PMC: 2020 Sep 1.
Published in final edited form as: Free Radic Biol Med. 2019 Jun 22;141:244–252. doi: 10.1016/j.freeradbiomed.2019.06.015

Figure 3. Tat Expressing Cells Exhibit Increased Oxidative Burden and Protein Modification.

Figure 3.

.(A) Total ROS is increased in HeLa-Tat-FLAG cells. Total ROS were measured by incubating cells with 1-Hydroxy-3-methoxycarbonyl-2,2,5,5-tetramethylpyrrolidine-HCl (CMH) and nitroxide was measured via EPR spectroscopy. Values shown as nmol of nitroxide per gram of protein.

(WT n = 6, TatFLAG n = 9) (***p ≤ 0.001 by un-paired student’s T-test)

(B) Mitochondrial ROS is increased in HeLa-Tat-FLAG Cells. Mitochondrial ROS were measured by incubating cells with (2-(2,2,6,6-Tetramethylpiperidin-1-oxyl-4-ylamino)-2-oxoethyl)triphenylphosphonium chloride (Mito-TEMPO-H) and nitroxide was measured via EPR spectroscopy. Values shown as nmol of nitroxide per gram protein. (n = 6) (*p ≤ 0.05 by un-paired student’s T-test)

(C) HeLa-Tat-FLAG cells demonstrate increased protein nitrosylation as measured by ELISA. Total 3-nitrotyrosine was measured by ELISA in HeLa-WT and HeLa-Tat-FLAG cells from total protein extract. (n = 6) (**p ≤ 0.01 by unpaired student’s T-test)