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. 2016 Feb 2;11(2):e0147978. doi: 10.1371/journal.pone.0147978

Fig 6. EOC-CM prevents SIRT1 post-translational modification and improved SIRT1 activity throughNOX4–NADPH oxidasedownregulation in rMC-1.

Fig 6

(a) SIRT1 activity in rMC-1 cultured for 24 hours by a fluorescent method. Cells in NG or HG conditions were exposed to NOX4siRNA in presence or not of db/m EOC-CM or db/db EOC-CM. Representative Western blots for NOX4 in rMC-1 total lysate cultured for 24 hours in HG. The efficiency of the NOX4small interfering RNA (100nM) was ~63%.NS = no significant difference. (b) SIRT1 immunoprecipitation in rMCs-1 lysates exposed to HG in presence of EOC-CM. Carbonylation was detected by first derivitizing the samples with DNPH and immunoblotting with DNP antibody. Nitration was detected by immunoblottingwith nitrotyrosine (NT) antibody.